The crate, to be present in large amounts in higher eukaryotes, including lower eukaryotes, including Dictyostelium discoideum and reptiles, mammals, and amphibians, have been discovered so far. Their function is stored among species indicating that it is essential to the function of eukaryotic cells is very as a composition and structure of the protein. The vRNA is the length of a species-specific, which varies between 86 and 141 bases. Further, a number of terms different from the biological organism vRNAs. Each 89 bases in length, 94 other: for example, 141 bases long frog mouse express vRNA and rats represent 2 vRNAs. hvg3 and hvg1, hvg2: people, representing the related vRNAs three. You are educated guess of the vRNA relatively long rodents and covered with more than one (R1) small vRNAs in other species. It is the difference of the type of vRNA but, vRNAs polymerase III promoters all elements are highly conserved. Each vRNAs is expected to be harvested in stem-loop structure similar.

Incidentally, the functional role of the vRNA is not important, it is considered to be a structural role. This conclusion, deterioration of secondary structure that does not affect the form of a vault that vRNA that is based on the observation, different types vRNAs are stored in well is expected. The function of vRNA Since opening in 1986, and vault, remains speculative. The several groups (most is located in the cytoplasm) suggesting a role of depot intracellular transport based on (R1, R3) the intracellular localization barrel, and its structure. The co-localization with cytoskeletal vault, suggesting that it may be transported by the cytoskeleton or cytoskeleton involved in maintenance or partially frames. In torpedo electric line, it is transported in nerve endings between the cell body to support the role of the cytoskeleton in desk transport has been shown to box. It is not known vRNAs arches and may or may not be involved in intracellular transport actually.

Multi-drug resistance (LRP) lung resistance-related protein (MDR)-associated protein is an (MRP). The MDR, is the main cause of failure of cancer treatment, it is associated with overexpression of MRP was found. This gene cDNA LRP encoding the cDNA was found to be the human major vault protein. Note that it is overexpressed in human tumor cell lines having many MDR phenotype was found to MVP / vault. It is possible to act by the vault to transport the drug from the (R1) its target.

with respect to functionality of the vRNA, vRNAs should be noted that it is connected to a human cancer cell line storage complex some it. People vRNAs three, must Vault of complex ratio does not reflect the expression level. Three vRNAs have different affinities for the (TEP1) telomerase-associated protein 1. The TEP1, be identical with any of the smaller bolt protein was found. The TEP1, components of the telomerase complex is evolutionarily conserved.

By generating a knockout model, an attempt to understand the role of the role in human disease frame and probably have been made. In the cellular slime mold, unlike mammals, there are three major protein safe. The destruction of two of these genes, suggesting a role in survival (R1), to prevent the proliferation and / or cellular proliferation. However, (including the MVP as a human being) mouse, the interruption of MVP, can lead to loss of the depot, there was no phenotype was observed. (Also lacks the vRNA of) TEP1 KO mouse phenotype conspicuous does not exist. Cancer cell lines showed overexpression of MVP, but compared to the wild-type, MVP KO mice did not show increased sensitivity to the drug. Interestingly, the increase in tumors induced in carcinogen VPARP deficient mice have been reported, vRNAs related to a particular disease in humans remains. If vRNAs is linked to a number of conditions subtly, with relevance and known frame together vRNA, the role of the mobile phone potential of the above, I would not be surprised it.

Without encoding the protein product of any of its array long viroids, there was confusion on how can induce the symptoms of plants. The evidence, I believe that RNA silencing is involved. First, changes in the viroid genome will be able to change its virulence dramatically. This reflects the fact that siRNA all generated that has a base pair additional less targeted RNA. Second, siRNA corresponding to the sequence of the viroid genome was isolated from infected plants. Finally, transgenic expression of non-infected hpRNA of potato spindle tuber viroid is, develop the symptoms of viroid well as all related. These data were reproduced in a double-stranded RNA intermediates, such as viruses, they show that after instructed by the enzyme Dicer, when it is cut into, siRNA is loaded into the RNA-induced silencing complex are. This is what causes the symptoms viroid classical containing sequences capable of complementing the pairing base to inhibit translation or induce degradation and messenger RNA own factory really viroids siRNA.

RNA has been identified as a sub-viral RNA, viral satellite RNA, viroids, human hepatitis delta virus and satellite (HDV) is a infectious pathogens known minimum. Encoding (HDAgs) viral proteins two HDV genome, the circular genome of single-stranded small, negative sense RNA containing a self-cleavable motif, and is configured open reading frame from (ORF). HDV requires a complex distribution of the surface antigen of hepatitis B virus (HBV), a virus. Small is replicated autonomously in the host plant and unencapsidated, single-stranded circular RNA encoding viroid is (~ 400 NT). Based on the presence or absence of (chloroplast and nuclear), viroids are classified into two families of the subcellular localization conserved regions and ribosome hammer. I own the code for a protein of genes required for the replication, satellite does not depend on the helper virus for its propagation. Both satellite and satellite virus nucleic acid contains the satellite. Satellite virus is composed of capsid single-stranded RNA genome by the protein encoded by satellite. Satellite nucleic acid comprising a satellite RNA of single-stranded RNA satellite subset of satellite RNA and double-stranded. Double-stranded RNA wrapped in a coat of satellite assistant viral proteins. (Also known as a satellite RNA viroid similar) the satellite RNA satellite RNA circular protein, a small linear encoding major satellite RNA: by encoding the protein capsid, single-stranded RNA genome, the three groups I have a satellite that is not classified.

I form the main part (ie, associated with viroid, vHDV, and RNA satellite RNA) of the database section of the one associated with the sub-viral RNA sequences. Their families, are indexed according to the species or genus in the classification system on the basis of criteria established by the International Commission on (ICTV) classification of the virus RNA mutants of all. Once appointed by the ICTV, certain types of use are classified based on sequence similarity it. If any, various agents, RNA, redundant thereof, sequence variants, and the number of size distribution of them is calculated based on the secondary structure of the exemplary embodiment, the family, designated by point database ICTV I choose to lead-point representation of the full name of the code of the type that is. For a particular type of each of the RNA, users can have access to a complete list of links to sequence identical to the sequence variants. Array of identical, might mean consistency improved fitness of in vivo for a particular option. Further, a link to a specific RNA motifs specific is shown in each mutant. From this point, it is possible to individually or in groups, to select the sequence, the user displays them in a different format using analysis software further to the user. In order to display in HTML format records these causes, it is dealing with subviral server directly using (see below) bioinformatics various tools, or possible, to facilitate the transfer to another program, the raw It is stored in a text of.

The several types of small RNA, have been proposed repressor of gene expression great potential for use in gene therapy. The (III of Pol) RNA polymerase III, little is located in the intracellular compartment of different intracellular small RNA expression products such as usual, RNA that is highly structured is ideal for some. We designed a cassette with the Pol III promoter of human cells for quality, high-level expression of the RNA of small nucleolus and nucleoplasm. Nuclear RNA small U6 (a snRNA), 5S ribosomal RNA (the rRNA), and destination intracellular and transcription levels were compared to express the transcription into components 7SL RNA of a signal recognition particle. RNA is a specific inhibitor, in this way, allowing these tapes can not necessarily predict the same input RNA in place in the cell some place the most effective. Small interfering RNA several insert (of siRNA) will be tested for efficacy. One of the inputs of the siRNA that reduces the expression of lamin when it is transcribed from the promoter of the U6 snRNA in the core when it is found in the nucleolus and cytoplasm and transfer of ink 5S rRNA’s and 7SL RNA-based It does not affect the expression of the laminate. In order to investigate the general nature of the driving U6 siRNA inhibition with siRNA targeting of HIV was further tested by co-transfection with provirus in cell culture. The degree of inhibition of HIV-1 varies between the insert, but the results, cassette U6 has been shown to allow expression of the siRNA-like inhibitors of HIV gene expression.

To the genomic DNA, and a gene of an organism, including leading to the production of specific proteins serve multiple functions specific to nucleic acid coding for protein synthesis. Transcription and translation: the fusion protein, two steps are included. Transcription creating extra copies of DNA sequences and RNA, translation is the subsequent processing used for the synthesis of RNA from the amino acids of the protein it really is. Production of proteins, inhibition of the transfer process, resulting in a blocking its function in the end. In order to understand the role better protein plays in a particular disease state, these compounds make a useful research tool, Sigma-Aldrich, offers a number of small molecules that inhibit protein translation.

RNA interference called (PTGS) post-transcriptional gene silencing (RNAi), typically by causing the destruction of mRNA specific molecules, RNA molecules are biological processes that inhibit gene expression. Historically, transcriptional gene silencing it is (PTGS) after co-suppression, it is known, such as suppression, the name of the other. Only after a thorough understanding of the process of apparently unrelated these, that they explained the phenomenon of RNAi all became clear it. 2006, Craig C. Mello and Andrew Fire, the publication is elegance in 1998 they shared the Nobel Prize in Physiology or Medicine for their work in RNA interference of nematode worm C,.

It is the center that the interfering RNA – two types of small ribonucleic acid (RNA) molecule – RNA and micro (the miRNA) small interfering RNA (the siRNA). The RNA, is a direct product of the gene, by preventing the mRNA from the protein production RNA and other specific, the (mRNA), for example, RNA small these, the molecules that increase or decrease its activity It can be coupled. Transposon and virus – not only plays an important role in protecting cells against parasitic nucleotide sequence, RNA interference, gene expression in general and to direct development.

The RNAi, located in eukaryotic organisms including many animals, the method, long double-stranded RNA enzyme dicer cutting (dsRNA) molecules into short fragments of 20 nucleotides difficult-dual, which is called siRNA of, It is initiated by the. In other words, each siRNA, has developed a direction and passenger strand of single-stranded 2 (SS) ssRNAs, you want to use. Passenger strand is reduced, the management direction contained RNA-induced silencing complex (RISC). Research results are post-transcriptional gene silencing that occurs when using a nucleotide sequence complementary to couple messenger RNA molecule chains and induce cleavage by catalytic component Argonaut, the RISC complex most. The restricted the molar concentration of siRNA Initially, in a particular organism, this process can spread throughout the body are known.

In cell culture, it is possible to induce suppression of specific genes of interest strongly selectively introducing synthetic dsRNA into cells, RNAi has become a valuable research tool for both in vivo. RNAi can be used for large screen to be canceled each gene to cells can be systematically and serve to identify the necessary components for event and cell specific processes, such as cell division. The road, it is used as a practical tool of medicine and biotechnology.

In eukaryotic cells, specific, and includes an enzyme having reverse transcriptase activity called telomerase. Telomerase is a reverse transcriptase which extends the ends of linear chromosomes. The synthesized “junk” DNA or DNA sequences that are repeated, telomerase RNA carry therefrom. This iterative sequence can be called a telomere DNA, is considered as a “cap” on chromosomes. Linear chromosomes since the duplicated each time, which is important, it is shortened. Not a repeat sequence is not essential for DNA sequence encoding a protein located at the ends of chromosomes, some of them, but ends of chromosomes, “hat” or “junk” DNA, the shortening is eliminated.

Often, without losing the DNA sequence cancer cells encode proteins important in order to be able to replicate their genome indefinitely, Telomerase is activated in cancer cells. Activation of telomerase may be part of a process that allows cancer cells become immortal. By using the remaining 10% of the alternate route is known as the alternate telomere elongation and maintenance ALT telomere, telomerase in recycling factor cancers indicates that occur in 90% of the carcinogenic all tumors in vivo by telomere elongation Because it is.

Found separated by Howard Temin at the University of Wisconsin-Madison in virion RSV, in David Baltimore independently, reverse transcriptase, was a 2 RNA tumor viruses at MIT in 1970 and R-MLV again RSV. So inconsistent with the central dogma of molecular biology said is transcribed into RNA, which is translated into protein DNA, the idea of ​​reverse transcription, it is very unpopular at first. However, in 1970, David Baltimore scientists and Howard Temin When is the possibility that the reverse transcription of both names independent reverse transcriptase, genetic information is allowed finally to be sent in this way I have found an enzyme involved. It is encoded by the viral reverse transcription using the enzymatic replication process, enzymes are used. Then, to use their genome the enzyme reverse transcriptase RNA in a DNA that is integrated into the host genome is replicated therewith, for example, reverse transcription of RNA viruses such as retroviruses. The reverse transcription, a DNA virus, such as hepadnaviruses such as, RNA can be used as a template for the assembly, and you can make the DNA strand. HIV-infected persons using this enzyme. It would not have been able to cause the failure to reproduce the Zu reverse transcription viral genome is incorporated into the host cell.

To create a double-stranded DNA and the transfer direction of transcription of the DNA product short initial RNA-dependent DNA synthesis to the acceptor template region at the other end of the genome that can be processed to reach by reverse transcriptase later The DNA-dependent DNA activity contains.  It is located at the 5 ‘end 3’ end of the retroviral RNA. Place called (PBS) primer binding sites are annealed primers viral RNA is called the leader of PBS FIG 5 ‘is referred to the end of RNA 3 end of the RNA to a site and, as a PBS U5’ have. It is developed with the double-stranded viral RNA primer tRNA is paired with form-based and PBS 22 and 14 nucleotides. Fact ‘reverse transcription Since DNA was synthesized from 3 that is located near the 5 PBS end of the primer in’ to 3 ‘(for RNA) 5 direction, by the end of the viral RNA, a rarity There is no ‘. Therefore, the position, the reverse transcriptase primer and achieves comprising (RN RNase H), and various enzymes DNA polymerase and several steps, ribonuclease H. Go to the 3 ‘end of the viral RNA, and it is required to develop a polynucleotide.  Reverse transcriptase of HIV have ribonuclease activity that degrades the viral RNA during the synthesis of DNA-dependent DNA polymerase and copy DNA with the sense DNA strand antisense DNA to form an intermediate double-stranded DNA virus (vDNA).

If you want to play in the life cycle of a retrovirus, and there are three different systems of the next. First, the present invention, reverse transcriptase to synthesize a complementary strand viral DNA and viral RNA, the DNA subsequently. The DNA polymerase of the host cell, when duplicating an integrated viral DNA, replication process of the second occurs. Finally, I can transcription of RNA polymerase II proviral DNA in RNA is packaged in virions. As a result, the mutation can occur in one or all duplicate one of these steps. Since then, that unlike the DNA polymerase of all the other does not have a high error rate on record at the time of the RNA correction capability, to DNA by reverse transcriptase. This high error rate can accumulate rapidly as compared with that mutation to examine the morphology of replication. In reverse transcriptase generation of commercially available, has been cited by the leadership as an error rate of up to 1 M-MLV 30,000 for base and base of 17,000 AMV from 1 by Promega. Also produce non-enzyme reverse transcription single nucleotide polymorphism, for example, to be involved in processes such as creating meaningless transcripts artificial transcription united and exon shuffling is shown. This reverse transcriptase that can be fully demonstrated in vivo, the switching operation of the template to be one reason for the discovery of transcripts without thousands annotate the genome of the model organism has been speculated was.

Important discoveries of many of biology, has emerged from (ribonucleic acid) study of RNA, including creative work in the field of structural biology biochemistry, genetics, microbiology, molecular biology, and molecular evolution . By 2010, scientists of 30, was awarded the experimental work, including the study of RNA, the Nobel Prize. Findings specific high biological significance, are described in this article. For additional information, please see the article on the history of the history of genetics and molecular biology. For more information, please refer to the article and nucleic acid RNA.

When we investigated the early first 1900s, differential chemical and biological between DNA and RNA is a clear, they are isolated, the name of the material from which the initial RNA “is “known as, DNA is” yeast nucleic acid nucleic acid with a pancreas “. “The ribose nucleic” two nucleic acids that contains the name of the common RNA, containing sugars using different diagnostic tests, chemistry, carbohydrate chemistry, indicating that it is. While the DNA is stable, the biochemical studies of other early, that (degeneration), was broken at high pH easily RNA in alkaline has been shown. For example, analysis of nucleoside composition that thymine nucleobases trace components, the number of dimethylguanine and a small amount of shoe dough lysine are included, and the RNA, it contains a nucleotide base, such as DNA uracil in place of this RNA I was shown.

Claims the result RNA that appeared in the late 1950s, leading to the synthesis of protein and thus, the formation of DNA, the concept of RNA is associated with a description of the jack of “central dogma of molecular biology” that. Complex genetic analysis of mutations in the lac operon early 1960s, E. coli, and instead of T4 RII bacteriophage, there is the genetic code and means for determining the nature of the RNA. MRNA of the cell population, short-term nature of the RNA of bacteria with nature very complex biochemical separation of mRNA challenging. Using the reticulocyte in vertebrates to produce a large amount of mRNA that is enriched (major protein bands hemoglobin 2) β globin RNA and encoding α very, this problem is overcome in 1960.

In the 1950s, the results of the experiment, the radioactive amino acid to label the rat liver “microsomes” very quickly after application it is associated with (redefined as ribosomes after) were found, before it will be is a cellular protein that was included widely been shown. The ribozyme, use an electron microscope, the ribonucleoprotein component thereof, biophysical methods capable of generating a (corresponding to hundreds of thousands of gravitational times) the acceleration is very high, ultracentrifugation sedimentation mainly To visualize that it has been identified by analysis. (Moves along the mRNA molecule number of ribosomes) polysomes It has been identified in the early 1960s, their research, generate processively proteins, ribosomal mRNA to the 5 ‘to 3’, as is the case they they do has led to the understanding of how to keep reading in the direction of.

Biochemical experiment painting, shows the radioactive amino acid is being built in a small RNA molecules remain soluble in the large RNA containing the conditions in which the particles to settle rapidly. These molecules will change the transfer RNA (of tRNA) (deer) and later referred to as water-soluble. That there is a set matching enzymes involved in the binding of (III) mRNA, the correct amino acids, subsequent studies, the plurality of specific amino acids, the mRNA species, each associated with (II) (I), each cell anticodon sequence of the tRNA to have has been shown to form a specific interaction with codons decoding mRNA.

The (RT-PCR), which is one of many embodiments of the polymerase chain reaction (PCR) Reverse transcription-polymerase chain reaction. This technique is used in the field of molecular biology for detecting the expression levels of RNA widely. RT-PCR has been confused real-time polymerase chain reaction and scholars and students (QPCR) often. However, they are different techniques clearly. By creating a (cDNA) the additional DNA from RNA transcription, RT-PCR, are used for the qualitative detection of gene expression but, QPCR was quantified amplification of DNA using the fluorescent probe . Quantitative PCR, quantitative real-time PCR, [3], QPCR also serves as a real-time quantitative PCR. Conventional PCR and RT-PCR, by amplifying the application of the two techniques in this way, to generate multiple copies of DNA isolation of specific, but is fundamentally different. Conventional PCR is used merely for the exponential amplification of DNA sequences. RT-PCR was used to using reverse transcriptase, cloning a gene that is expressed by reverse transcription of RNA of interest in the complement of the DNA. The cDNA then was newly synthesized was amplified using conventional PCR.

In addition to the qualitative study of gene expression by including an equivalent QPCR, RT-PCR may be used to quantify the RNA absolutely and relatively. Often, combined technique described by (real-time quantitative RT-PCR sometimes) Quantitative RT-PCR, real time RT-PCR is an abbreviated RRT-PCR QRT-PCR, or RT-QPCR,. Most powerful compared to other methods for quantitative RNA by Northern blotting, detecting the level of the QRT-PCR RNA, can be considered quantitative analysis with high sensitivity. Infection and expression of one or more genes and this is used to analyze the expression patterns for the identification of disease often.

Quantification of mRNA using RT-PCR may be achieved either two-step reaction or one step. Difference between the two approaches is the number of tubes used in the course of the procedure. The approach of one stage, all of the cDNA synthesis reaction by PCR amplification occurs in the tube. On the other hand, require a reverse transcriptase reaction, the two-stage reaction was performed in a separate tube PCR amplification. The one-step method, I thought to minimize the deviation from the experiment of the enzyme reaction of all in the environment. However, it is susceptible to degradation in the approach of a single step, and the use of this approach, starting template RNA does not recommend that you repeated analysis of the same sample. Furthermore, the one-step approach, it is less accurate than a two-step approach was reported. The main advantage of the two-step approach, analysis with high reproducibility shows a correlation coefficient in the range of 0988-0974. Removal of primer dimers, so can be achieved by simple modification of the melting temperature, for example, be used DNA binding dye SYBR Green or the like, it is the preferred method of analysis. A disadvantage of two-stage approach is sensitive to contamination by more frequent handling samples.

Serum prostate-specific antigen (PSA) assay has a high false positive rate. Marker PSA is to provide diagnostic information is limited. As well as tumor, multi-marker test for detection of potentially fatal, provides a clinical need that is not met. Using the nanoString nCounter system, 20 – as a means for detecting prostate cancer, gene expression of the gene multiple test was developed based on the digital count gene RNA transcripts urine. In this test, centrifuged to pellet void urine, cells were amplified preselected hybridization probe sets the RNA was purified. Measured by DNA microarray, amplification test cell line RNA is not a gene rich level and the number of matches and bias considerable, but will be displayed. For data analysis, each number, to compare β2 microglobulin, and gene family. I have analyzed urine samples of five preoperative patients, and 2 non-cancer. Pathology information is restored. The low June 06 and Gleason scores and prostate cancer markers noncancerous low, are known, the signal for the majority of genes, were positive when. In contrast, one instance Gleason 4 Figure 5 is a strong signal for all associated with cancer markers including CD24. The non-cancer, there is shown a signal of cancer marker of 6 all, there is a possibility that undiagnosed, harbor cancer this person. Analyze the natural wastes, this multiplex assay can be used to stratify patients and early detection potential screening of cancer. The diagnostic information, I got from RNA signature associated with the cells of prostate cancer.

It is a mechanism in prokaryotes that causes the RNA transcription (referred not to Rho-dependent termination also) internal termination is stopped. In this mechanism, mRNA is cytosine – Includes an array can be calculated in a pair with itself to form a stem-loop structure 7-20 base pairs in length guanine base pair is rich. These bases, so to form three hydrogen bonds with each other, very strong. I have a chain of uracil residues after the stem-loop structure. Link between the uracil and adenine is very weak. Polymerase closely enough to bind to stem loop structure To be thrown down temporarily proteins associated with RNA polymerase (Nusa Dua). This break, poly – are consistent with transcription polymerase sequence of uracil. Uracil bonds – weak adenine can be dissociated to relax from the RNA polymerase, lowering the energy of the destabilization of the double-stranded RNA-DNA.

Will cause the RNA polymerase to stem-loop structure that do not comply by poly uracil sequence will pause, but so is stable too to fully relax to cause an end to the double, usually, it after a short time, I will continue to transcription.
In many cases, ρ-independent transcription termination mechanism is the basis of the activity of the RNA cis-acting regulatory elements, as riboswitch.

E. coli genome equencing whole may have a termination sites most of the operon is not dependent on the row are shown. In the transcribed RNA, a series of U residues in front of those of self-complementary region a GC-rich and nucleotides intervening some: I have two characteristics in extreme sequence of these. Area of ​​the self-growth of additional RNA strand are synthesized with each other to form a stem-loop structure, base pairs of additional sequences. The stem-loop structure is believed to interact with the recording of E. coli RNA polymerase and cause it to break. Further, base pairs between the residues in the template DNA strand U and Watson 3 ‘end of the residual nascent RNA chain – very unstable as compared to other types of Crick base pairs. These two features, have solved the release of the RNA strand of the transcription complex at the end sites that do not depend on Rho probably. Mutations that decrease and results dyad symmetry to U, a continuous low end to reduce the part of these sites.

Several mechanisms for the regulation of transcription is found in eukaryotes and bacteria. RNA polymerase plays a role of two primary mechanisms for the conclusion of the occurrence transcription in E. coli. Additional proteins, called rows, transcription terminators, a mechanism is required, but not the other. In many cases, these mechanisms are referred to as dependent end of Rho If you do not depend on low. In eukaryotes, the mechanism of transcription termination seems to be different in each RNA polymerase three. Here, I focus on the end of the first mechanism in bacteria that will be understood from the mechanisms of eukaryotic organisms has been described by way of example research well two of termination of transcription of eukaryotic more clearly.

Another mechanism for the regulation of gene expression in bacteria, called recognized by the analysis of a particular strain of E. coli mutant first tryptophan operon is attenuated. In the presence of tryptophan, tryptophan repressor, please recall that to inhibit the transcription of TRP operon. However, it is not exhaustive, according to protein synthesis, this inhibition is complemented by the weight loss to provide the level of regulation of tryptophan synthesis.

In essence, attenuation results in early termination of transcription at the attenuator site located downstream of the coding sequence of the enzyme TRP and before, from the operator region – promoter. The presence of E. coli, the tryptophan and mutant with a small deletion in the area of ​​manufacturing tryptophan operon more than the normal amount of tryptophan synthase in the absence. Trp mRNA of the Trp promoter nearby sequence comparison of researchers at the start of the RNA molecule, deletions and these, or may lead to increased synthesis of DNA region of Trp in both mutant and normal E. mRNA molecules of normal, contains 162 nucleotides upstream of Trp August the first initiation codon of the enzyme that is encoded TRP-five in order to understand. This part of the mRNA are referred to as (L) leader sequence. MRNA transcript tryptophan rich TRP only and will not be transferred consists of the rest of the 7 KB TRP operon ≈ and only leader RNA by virtue of the action of the tryptophan repressor of E. coli cells normal production the transcription of these. When tryptophan is limited, however, comprises a coding sequence for all of the enzyme is encoded trp promoter and leader sequence, tryptophan operon is transferred as a whole. normal cells than do mutant cells is the attenuation region has been deleted, to produce a mRNA of Trp twist in all conditions.

Growth rate is a step-by-step addition of amino acids to growing protein chain. The order of the amino acid, I was determined from an array of codons of mRNA in
. Please refer to the “Genetic code expansion process forward in order eEFs is necessary in order to facilitate the expansion factor protein, the cycle (eukaryotic elongation factor) following.:
1. It has been picked up by extension EEF-1 and in the presence of amino group-containing mRNA molecule of GTP (aminoacyl tRNA AA-tRNA is).
2. Complex result of entering the A site empty of peptidyl-tRNA or Met-tRNAi initiator of carrying ribosome.
3. It is matched with mRNA codon ribosome to place at the scene anticodon of the incoming aminoacyl tRNA. It can be arranged in different combinations of translator 64 must be able to select a supported aminoacyl tRNA anticodon the three base codon is matched. In this proof-reading, in order to aminoacyl-tRNA and the anticodon of non-related thrown out from the ribosome, you want to check, and replace the AA-tRNA’s new.
4. By peptide bonds, and is associated with new amino acid at the scene almost immediately after the right aminoacyl-tRNA enters the site of growth peptide P-site. Peptide bond formation catalyzed by the ribosome itself. The reaction was left of peptidyl-tRNA and new ribosomal P-site of the mRNA of the blank in the site.
5. One codon of the mRNA is moved forward in the next step of the ribosome. At the same time, the shift of the mRNA empty, it will be forwarded to the P-site from the field from the P site of the E-site as the mRNA of peptidyl. Process is facilitated by GTP and elongation EEF-2.
6. Located in the rolling after P-site of the peptidyl-tRNA, the next codon of the mRNA is available for reaction with the aminaoacyl-tRNA another site.
7. The ribosome, these reaction steps are repeated until it encounters the frame stop codon. At this point, the transfer is completed.

As stage of regulation, considerable progress has identified this stage and highly dynamic transcription cycle to extend the transcript of RNA polymerase II (RNAP II). In this section, we discuss the many factors that regulate the elongation phase of transcription. Elongation factor classic that modulate the activity of RNAP II still, we include certain factors that facilitate the expansion of the chromatin template in recent years and more. In addition, we discuss the factors to be associated with RNAP II, but it does not want to regulate their catalytic activity. Extension, has emerged as a process of coordination required multiple stages of maturation and biosynthesis of mRNA.

And complete 12 subunit RNA polymerase that is associated with the product (POL) II transcription bubble, the crystal structure of the RNA template, non-template DNA 7 base pair DNA / RNA hybrid, three in each of the separation of DNA and RNA incoming I discloses the nucleotide of one. Complex accepts posttranslocation state, houses the (NTP) substrate triphosphate co-crystal nucleoside. The binding to the active site of the pores that can interact with DNA-based matrix in the NTP. Residue of NTP around, are reserved and suggest a universal mechanism for inclusion and NTP selection, the RNA polymerase of all cells. DNA-RNA separation circuit and DNA-DNA may be explained by the failure of the duplex Pol II induced. That contributes to the integration of hybrid, to prevent chain recombination, and the share of four protein chains of active center cleft creates the exit tunnel of RNA. Binding of TFIIS elongation aligned RNA of the active center, converting a complex growth to another state where necessary, difficult to blocking.

Regulation of gene expression, is one of research areas most intensively in the whole of science. It is based on the cell type specific gene expression of different multicellular organism. Deregulation of the appropriate pattern of gene expression, a significant effect on the basis of many diseases of cell function. There are gene expression, and many cellular processes that control the most direct regulation during transfer. Transcription of the gene encoding the protein in eukaryotes is performed by RNA polymerase II (RNAP II). Until recently, most studies aimed to reveal the molecular mechanism of transcriptional control is focused on the early stages including creating (see below) to the start set of (before) the transcription start like this. Over the years, transcription elongation is considered as a trivial addition of ribonucleoside triphosphates into mRNA chain growth. This, it is a regulatory step highly dynamic and transcription cycle that can this process to adjust the downstream event is immediately obvious. Many factors that targeting the elongation phase of transcription in particular has been confirmed. Importantly, multiple steps, is modulated by the interaction transcription elongation complex of RNAP II with (TEC) including exports for pre-mRNA splicing ceiling, 3′-end processing, and supervision, and mature mRNA.

Many changes, it is necessary so that it extends from the transition to transcription initiation occurs in RNA polymerase II (Pol II) transcription complex. In the intermediate stage of the transfer, the trigger contact factors are lost and stable relationship with nascent transcripts has been established. These include collective promoter clearance is included. After transcription elongation complex, the property has reached a point that could not be distinguished from those of the composite transcript promoter clearance much longer is complete. The distance method of Pol II, is composed of a series of steps, the distance downstream surprisingly long transfer extends to. It is part of a Special Issue, and this article is entitled: transcription elongation of RNA polymerase II.

By monitoring stretched mechanically rolling, the extended end-to-end single DNA molecule, the RNA polymerase (RNAP), to monitor the change of the extension corresponding to the arrangement around the turn of the promoter DNA directly we I was able. By carrying out the experiments in parallel with winding DNA positively and negatively, we extended to be due to (RNAP-dependent DNA compaction and (at ≈ 1-BP resolution) changes the extension caused by RNAP-dependent DNA evolution with a resolution of ≈ 5nm was able to deconvolution the change). For the quantification of the effects supercoiled and dynamics, the degree of development and integration, development, of the dense, sequence, the nucleotide ppGpp Noto, we were using this approach. Further, in order to detect the recycling of promoter and promoter clearance by a succession molecules RNAP, we were using this approach. We (by the number and position of the coiled-coil) exercise super coil structure (rather than through the torque) the mechanical effects believe that stable rate and formation of advanced composite body as being dependent deeply in super coil. This approach should allow the analysis of nucleic acid processing and other factors that cause changes in DNA compaction and / or twist of DNA.

Comprising the reaction sequence initiation of transcription,: (I), the promoter DNA around in order to bind to the promoter DNA to form the RNAP promoter gated, to form (II) RNAP RNA polymerase holoenzim is (RNAP) (III) RNAP avoid promoter (in a process called “promoter clearance”) and, (referred to as “promoter relaxation”) RNAP open promoter complex that generates one rotation, the RNA productive by elongation complex of RNAP-DNA I go into the synthesis.

We have developed the approach of the DNA molecule nano manipulation that we can develop the promoter by RNAP, to detect the promoter well-formed, characterized. Our approach is using the experimental apparatus was developed for the DNA polymer physics originally. In this experiment, the multiple links paramagnetic beads multiple links to the glass surface is connected at one end to the other end double-stranded DNA molecules containing a promoter region, DNA is limited mechanically has been, by applying a pair of magnets in the helical axis of the tension DNA twisting, between the glass surface and the neck (corresponding to the extension of the DNA end-to-end, L) surface of the glass and the head of the The distance between can be observed in real time using a video microscope. A pair of magnets is rotated, the supercoiled off rotation is provided a lock-step on the neck, was introduced into the torsion limit DNA molecule registry lockstep, which is supercoiling, therefore, L is changed . In this experiment, it is possible to construct a calibration curve experimental rotational speed of counterclockwise relates superhelical turns or negative or positive clockwise pair of L magnet thus is easy. Over a wide range of supercoiled positive and negative changes in L of 56 ± 5 nanometers supercoiled turn, under the conditions of our and l varies with the number of turns of the coiled-coil of the positive or negative linearly.

The first step of transcription, is an opening that identifies the start of the mRNA position and appropriate it. Due to the nature of the “out under” genetic code, the location of the start codon only, has the potential to determine the reading frame, false detection can lead to a complete distortion of messages as a result of a change in reading frame. Exact start is essential for an accurate translation. The genetic code, the start of a particular signal is not included. Rather, the dual function that is associated with the presence of two types of mRNA codons August methionine. One, met for use in normal-tRNAMets initiator tRNA and especially at first. Met-tRNAMet second, are used amino acid methionine at the position of the inside, in the process of expansion of the set, including reasonable-elongator only protein. (Initiation factor EIF of eukaryotic) initiation factor of several proteins that provide support to the ribosome during the initiation process.
When starting the process, it may be divided into separate steps several.
1. First, ribosome dissociation is called the 60’s and 40’s department in the configuration sub-unit of the two. Subunit can prevent spontaneous reassociation of connecting the initiation factor EIF-3 in the ribosomal small subunit.
2. Initiation factor 2 is an EIF-GTP-binding protein that recognizes a specific initiator tRNA to form a ternary complex and (A).
3. Ternary complex B is associated with the 40S ribosomal subunit.
4. 5 ‘end of the RNA cap structure is recognized by initiation factor EIF-4F. This factor, interacts with the cap structure in particular: Has division (see cap structure splicing). Combination of the user who started the cause of activation is 40S, subunit initiator tRNA, including the 5 ‘end of the mRNA. This type of start, is known as a cap-dependent start, which is the most common type of start. May be caused by a cap start mechanism independent of another start.
5. Move the 5 ‘untranslated end of the mRNA of the related subunit mRNA in until it reaches the codon of the first of August to become the start codon of the translation process. It requires energy (ATP), the process called scanning is started additional factors.
6. Subunit codons 60 begins to bind to the 40S subunit when it reaches the activation of 40S subunits. This reaction, to release the factors that start from the ribosome as this requires factor EIF-5 start of hydrolysis add EIF-2 bound GTP. As a result of the start process, the initiator will be placed in the collapse of the ribosomal P-80S ribosome site.The of 60S subunit and 40S. Subunit can prevent reassociation spontaneous intiation factor EIF-3.

The basic process of protein production, the addition of amino acids one by one to the end of the protein. This operation is performed by the ribosome. Selection of the type of amino acid to be added is determined by the mRNA molecule. Each amino acid, I was added to match the base sequence of the mRNA of the three. Type of amino acid of one particular possible is accepted only in each triplet such. Contiguous amino acids are added to the adaptive circuit to the nucleotide triplet continuous mRNA. As a result, I will determine the sequence of amino acids of the chain of amino acids nucleotide sequence of the mRNA template of the chain has been formed. It is found in the C-terminus of the peptide, and thus, the addition of amino acids, the transfer function – is said to be a carboxyl – amino.

carry the genetic information encoded as a ribonucleotide sequence of chromosome of the ribosome during mRNA. By translational machinery in the sequence of nucleotide triplets called codons ribonucleotides are “read”. Each triplet of these codes for a particular amino acid. Molecule of the ribosome translates the code to this specific sequence of amino acids. The ribosome is a multi-subunit structure containing the protein and ribosomal RNA. This is the “factory” is incorporated amino acid, the protein. (93 nucleotides from 74) small non-coding RNA chain, amino acids are transported ribosome mRNA. Of tRNA is, have the site of a site called anticodon and amino acid attachment. The RNA, to be complementary to RNA triplet, encoding the amino acid of the cargo in the anticodon triplet.

Aminoacyl-tRNA synthetase (enzyme) catalyzes the coupling between amino acids, that the anticodon sequence tRNA and requires a specific. The product of this reaction is an aminoacyl tRNA molecule. This aminoacyl-tRNA, codon of the mRNA moves to the ribosome that is matched to the base of the pair added to the tRNA anticodon specific. The ribosome has three sites for tRNA to bind. They are (E for short) exit point aminoacyl site (abbreviation), and peptidyl site (abbreviated P). ‘For mRNA for moving to end, 3 seats, EPA 5’ 3 of the mRNA is 3 ‘orientation from the ribosome. I will bind and tRNA codon additional site of mRNA is imminent. I will hold the polypeptide chain mRNA of P management is growing. TRNA in the E site does not have the amino acid.

Initially, when bound to the codon that corresponds to that of the mRNA, are on the site aminoacyl-tRNA. Thereafter, the amino acid sequence of the mRNA and form of the peptide bond between amino acids mRNA to one location is transmitted to the site of mRNA growth and loaded polypeptide chain on the P site. Transfer occurs, and amino acids to E site that was in the accused now site at polypeptide chain, the tRNA, the P, move the mRNA leave the E site to a site that you are in the site P without the tRNA is now , aminoacyl-tRNA other enters the site to repeat the process. After being added to the circuit a new amino acid, the energy provided by hydrolysis of GTP which is connected to EEF-2 and (prokaryotes) translocase EF-G, the termination codon ribosome at the 3 ‘end of (eukaryotic) I want to move down. Energy necessary for translation of proteins is important. For N protein-containing amino acids, the number of high-energy phosphate bond necessary for translation 4N-1. Translation rate is different, but it is significantly higher (up to 17-21 amino acid residues in the second) prokaryotic cells than (amino acid residues within 6-9 pieces second) eukaryotic cells.