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1.
Codon choice in genes depends on flanking sequence information—implications for theoretical reverse translation 下载免费PDF全文
Karthikeyan Sivaraman AswinSaiNarain Seshasayee Patrick M. Tarwater Alexander M. Cole 《Nucleic acids research》2008,36(3):e16
Algorithms for theoretical reverse translation have direct applications in degenerate PCR. The conventional practice is to create several degenerate primers each of which variably encode the peptide region of interest. In the current work, for each codon we have analyzed the flanking residues in proteins and determined their influence on codon choice. From this, we created a method for theoretical reverse translation that includes information from flanking residues of the protein in question. Our method, named the neighbor correlation method (NCM) and its enhancement, the consensus-NCM (c-NCM) performed significantly better than the conventional codon-usage statistic method (CSM). Using the methods NCM and c-NCM, we were able to increase the average sequence identity from 77% up to 81%. Furthermore, we revealed a significant increase in coverage, at 80% identity, from < 20% (CSM) to > 75% (c-NCM). The algorithms, their applications and implications are discussed herein. 相似文献
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Robin Craig 《Journal of theoretical biology》1981,88(4):757-760
It is shown on a theoretical basis that the existence of a “power law” relationship between body mass M and total metabolic heat generation rate Q of the form Q = kMα does not uniquely determine the dependence of metabolic rate on body temperature. However, it is shown that a particular assumption for this temperature dependence, successful in other problems, does predict a “power law” similar to the empirical one. At the same time it also accounts satisfactorily for the linear dependence of metabolic rate on ambient temperature. 相似文献
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Codon usage can affect efficiency of translation of genes in Escherichia coli. 总被引:45,自引:19,他引:26 下载免费PDF全文
M Robinson R Lilley S Little J S Emtage G Yarranton P Stephens A Millican M Eaton G Humphreys 《Nucleic acids research》1984,12(17):6663-6671
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An Arabidopsis thaliana T-DNA mutagenized population (GABI-Kat) for flanking sequence tag-based reverse genetics 总被引:16,自引:0,他引:16
The GABI-Kat population of T-DNA mutagenized Arabidopsis thaliana lines with sequence-characterized insertion sites is used extensively for efficient progress in plant functional genomics. Here we provide details about the establishment of the material, demonstrate the population's functionality and discuss results from quality control studies. T-DNA insertion mutants of the accession Columbia (Col-0) were created by Agrobacterium tumefaciens-mediated transformation. To allow selection of transformed plants under greenhouse conditions, a sulfadiazine resistance marker was employed. DNA from leaves of T1 plants was extracted and used as a template for PCR-based amplification of DNA fragments spanning insertion site borders. After sequencing, the data were placed in a flanking sequence tag (FST) database describing which mutant allele was present in which line. Analysis of the distribution of T-DNA insertions revealed a clear bias towards intergenic regions. Insertion sites appeared more frequent in regions in front of the ATG and after STOP codons of predicted genes. Segregation analysis for sulfadiazine resistance showed that 62% of the transformants contain an insertion at only one genetic locus. In quality control studies with gene-specific primers in combination with T-DNA primers, 76% of insertions could be confirmed. Finally, the functionality of the GABI-Kat population was demonstrated by exemplary confirmation of several new transparent testa alleles, as well as a number of other mutants, which were identified on the basis of the FST data. 相似文献
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Selective translation of heat shock mRNA in Drosophila melanogaster depends on sequence information in the leader. 总被引:26,自引:3,他引:26 下载免费PDF全文
One of the effects of a temperature increase above 35 degrees C on Drosophila melanogaster is a rapid switch in selectivity of the translational apparatus. Protein synthesis from normal, but not from heat shock, mRNA is much reduced. Efficient translation at high temperature might be a result of the primary sequence of heat shock genes. Alternatively a mRNA modification mechanism, altered as a consequence of heat shock, might allow for efficient high temperature translation of any mRNA synthesized during a heat shock. The gene for alcohol dehydrogenase (Adh) was fused to the controlling elements of a heat shock protein 70 (hsp70) gene. Authentic Adh mRNA, synthesized from this fusion gene at elevated temperatures was not translated during heat shock. A second Adh fusion gene in which the mRNA synthesized contained the first 95 nucleotides of the Hsp70 non-translated leader sequence gave rise, at high temperature, to mRNA which was translated during the heat shock. Thus, the signal(s) in the mRNAs controlling translation efficiency at heat shock temperatures is encoded within the heat shock genes. 相似文献
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Codon usage determines translation rate in Escherichia coli 总被引:42,自引:0,他引:42
We wish to determine whether differences in translation rate are correlated with differences in codon usage or with differences in mRNA secondary structure. We therefore inserted a small DNA fragment in the lacZ gene either directly or flanked by a few frame-shifting bases, leaving the reading frame of the lacZ gene unchanged. The fragment was chosen to have "infrequent" codons in one reading frame and "common" codons in the other. The insert in these constructs does not seem to give mRNAs that are able to form extensive secondary structures. The translation time for these modified lacZ mRNAs was measured with a reproducibility better than plus or minus one second. We found that the mRNA with infrequent codons inserted has an approximately three-seconds longer translation time than the one with common codons. In another set of experiments we constructed two almost identical lacZ genes in which the lacZ mRNAs have the potential to generate stem structures with stabilities of about -75 kcal/mol. In this way we could investigate the influence of mRNA structure on translation rate. This type of modified gene was generated in two reading frames with either common or infrequent codons similar to our first experiments. We find that the yield of protein from these mRNAs is reduced, probably due to the action in vivo of an RNase. Nevertheless, the data do not indicate that there is any effect of mRNA secondary structure on translation rate. In contrast, our data persuade us that there is a difference in translation rate between infrequent codons and common codons that is of the order of sixfold. 相似文献
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N D Cook 《Journal of theoretical biology》1977,64(1):113-135
The theoretical and experimental evidence for a “reverse translation” mechanism in animal cells is reviewed. Mekler's (1967) theory is presented as the most likely means for reverse translation. This theory is shown to be consistent with the postulate of the “central dogma” that molecular information does not pass out of protein molecules once it has gotten in.The importance of nucleic acid changes in: (1) the immune response, (2) evolution, (3) cancer, (4) cell differentiation, and (5) learning in animal brains is mentioned and each topic is related to the reverse translation hypothesis. Because of the intense research effort in immunology, the experimental data which indicate an active role for antigen in antibody formation and the data indicating the importance of RNA in the immune response are dealt with more thoroughly. 相似文献
12.
We have examined codon bias in 207 plant gene sequences collected from Genbank and the literature. When this sample was further divided into 53 monocot and 154 dicot genes, the pattern of relative use of synonymous codons was shown to differ between these taxonomic groups, primarily in the use of G + C in the degenerate third base. Maize and soybean codon bias were examined separately and followed the monocot and dicot codon usage patterns respectively. Codon preference in ribulose 1,5 bisphosphate and chlorophyll a/b binding protein, two of the most abundant proteins in leaves was investigated. These highly expressed are more restricted in their codon usage than plant genes in general. 相似文献
13.
Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes 总被引:665,自引:0,他引:665
By analyzing the effects of single base substitutions around the ATG initiator codon in a cloned preproinsulin gene, I have identified ACCATGG as the optimal sequence for initiation by eukaryotic ribosomes. Mutations within that sequence modulate the yield of proinsulin over a 20-fold range. A purine in position -3 (i.e., 3 nucleotides upstream from the ATG codon) has a dominant effect; when a pyrimidine replaces the purine in position -3, translation becomes more sensitive to changes in positions -1, -2, and +4. Single base substitutions around an upstream, out-of-frame ATG codon affect the efficiency with which it acts as a barrier to initiating at the downstream start site for preproinsulin. The optimal sequence for initiation defined by mutagenesis is identical to the consensus sequence that emerged previously from surveys of translational start sites in eukaryotic mRNAs. The mechanism by which nucleotides flanking the ATG codon might exert their effect is discussed. 相似文献
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SiteFinding-PCR is a method for isolating flanking sequence tags (FSTs) of T-DNA insertion lines, but the efficiency needs to be improved. Here we report a computation-assisted design for the random primers used in SiteFinding- PCR. A short sequence, GCATG, was screened from the rice genome and used as the 3' end of the random primer. When applying the optimized primer for isolating FSTs from 168 transgenic rice lines, we obtained 107 specific products, including 64 FSTs. The efficiency of obtaining FSTs using the modified version of SiteFinding-PCR increased by 73.0% compared with the method previously reported (P < 0.01, μ test). We also provide computational results for several other plant species such as maize, sorghum, Arabidopsis, foxtail millet, and Brachypodium based on the available genome data, so that the modified method could be easily adapted to other species. 相似文献
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Codon contexts in enterobacterial and coliphage genes 总被引:6,自引:0,他引:6
This investigation of the codon context of enterobacteria, plasmid, and
phage protein genes was based on a search for correlations between the
presence of one base type at codon position III and the presence of another
base type at some other position in adjacent codons. Enterobacterial genes
were compared with eukaryotic sequences for codon context effects. In
enterobacterial genes, base usage at codon position III is correlated with
the third position of the upstream adjacent codon and with all three
positions of the downstream codon. Plasmid genes are free of context
biases. Phage genes are heterogeneous: MS2 codons have no biased context,
whereas lambda genes partly follow the trends of the host bacterium, and T7
genes have biased codon contexts that differ from those of the host. It has
been reported that two successive third-codon positions tend to be occupied
by two purines or two pyrimidines in Escherichia coli genes of low
expression level. Here, the extent to which highly expressed protein genes
can modulate base usage at two successive codon positions III, given the
constraints on codon usage and protein sequence that act on them, was
quantified. This demonstrates that the above-mentioned favored patterns are
not a characteristic of weakly expressed genes but occur in all genes in
which codon context can vary appreciably. The correlation between
successive third-codon positions is a distinct feature of enterobacteria
and of some phages, one that may result from adaptation of gene structure
to translational efficiency. Conversely, codon context in yeast and human
genes is biased--but for reasons unrelated to translation.
相似文献
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Patterns in codon usage were examined for the coding regions of the 23 known lepidopteran hemolymph proteins. Coding triplets
are GC rich at the third position and a significant linear relationship between GC content of silent and nonsilent (replacement)
sites was demonstrated. Intron GC content was significantly lower than in coding regions and no relationship between intron
GC content and the same at silent and nonsilent sites was found. Though hemolymph proteins are all produced by the same tissue—fat
body—significantly less bias was observed when all moth sequences were pooled than when sequences of the two major species
were analyzed separately, as predicted by the genome hypothesis. In cases where no statistically significant bias was observed,
polar or acidic basic amino acids were almost exclusively involved. Calculation of codon adaptation indices (CAI) was of limited
value in quantifying the degree of codon bias and probably reflects the complexity of multicellular-organism life cycles and
the changing patterns of gene expression over different developmental stages.
Correspondence to: D.R. Frohlich 相似文献
19.
We found that the peptide Gly-Arg-Gly-Glu-Ser-Pro (GRGESP) inhibited spreading of human fibroblasts inside collagen gels and markedly decreased gel contraction, but this peptide had no effect on cell spreading on collagen-coated surfaces. On the other hand, the peptide Gly-Arg-Gly-Asp-Ser-Pro (GRGDSP), which inhibited cell spreading on collagen-coated surfaces, did not inhibit cell spreading within collagen gels and was a less effective inhibitor of collagen gel contraction than GRGESP. Based on these findings, we conclude that human fibroblasts can interact with different collagen cell recognition sequences depending upon topographical organization of the collagen. 相似文献
20.
Codon usage in Plasmodium vivax nuclear genes was analysed and compared with that in Plasmodium falciparum nuclear genes. Preferred codons were determined for P. vivax. Unlike P. falciparum, P. vivax genes are about 15% less A+T rich in the coding regions, with no obvious A+T bias at the third position of the codons. The amino-acid composition of P. vivax gene products is also different from that of P. falciparum. These results provide valuable information to facilitate gene cloning as well as expression and transfection studies for P. vivax. 相似文献