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The aim of this study was to analyze patterns of nucleotidic composition and codon usage in the pea aphid genome (Acyrthosiphon pisum). A collection of 60,000 expressed sequence tags (ESTs) in the pea aphid has been used to automatically reconstruct 5809
coding sequences (CDSs), based on similarity with known proteins and on coding style recognition. Reconstructions were manually
checked for ribosomal proteins, leading to tentatively reconstruct the nea-complete set of this category. Pea aphid coding
sequences showed a shift toward AT (especially at the third codon position) compared to drosophila homologues. Genes with
a putative high level of expression (ribosomal and other genes with high EST support) remained more GC3-rich and had a distinct
codon usage from bulk sequences: they exhibited a preference for C-ending codons and CGT (for arginine), which thus appeared
optimal for translation. However, the discrimination was not as strong as in drosophila, suggesting a reduced degree of translational
selection. The space of variation in codon usage for A. pisum appeared to be larger than in drosophila, with a substantial fraction of genes that remained GC3-rich. Some of those (in
particular some structural proteins) also showed high levels of codon bias and a very strong preference for C-ending codons,
which could be explained either by strong translational selection or by other mechanisms. Finally, genomic traces were analyzed
to build 206 fragments containing a full CDS, which allowed studying the correlations between GC contents of coding and those
of noncoding (flanking and introns) sequences. 相似文献
3.
Mikael Falconnet 《Mathematical biosciences》2010,224(2):101-108
We consider models of nucleotidic substitution processes where the rate of substitution at a given site depends on the state of the neighbours of the site. We first estimate the time elapsed between an ancestral sequence at stationarity and a present sequence. Second, assuming that two sequences are issued from a common ancestral sequence at stationarity, we estimate the time since divergence. In the simplest non-trivial case of a Jukes-Cantor model with CpG influence, we provide and justify mathematically consistent estimators in these two settings. We also provide asymptotic confidence intervals, valid for nucleotidic sequences of finite length, and we compute explicit formulas for the estimators and for their confidence intervals. In the general case of an RN model with YpR influence, we extend these results under a proviso, namely that the equation defining the estimator has a unique solution. 相似文献
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S E Klebanoiv Iu M Bochina A L Berman 《Zhurnal evoliutsionno? biokhimii i fiziologii》1991,27(3):289-294
The method of cDNA sequences comparison is suggested. The method consists of constructing the dot matrixes for each codon position separately. The fastest disappearance of the information about relationship is observed when the third nucleotides in codon are compared, and the comparison of the second nucleotides in codon turned out to be effective so as the comparison under acids. 相似文献
6.
The objective of this work was to estimate the nucleotidic variation between two groups of tepezcuintles (Agouti paca) from the states of Campeche and Quintana Roo, Mexico and within members of each group. Blood samples were collected from eleven A. paca kept in captivity. DNA from leukocytic cells was used for Ramdom Amplification of DNA Polimorphism (RAPD). The primers three 5'-d(GTAGACCCGT)- 3' and six 5'-d(CCCGTCAGCA)- 3' were selected from de Amersham kit (Ready.To.Go. RAPD Analysis Beads, Amersham Pharmacia Biotech), because they produced an adequate number of bands. The electrophoretic pattern of bands obtained was analyzed using software for phylogenetic analysis based on the UPGMA method, to estimate the units of nucleotidic variation. The phylogenetic tree obtained with primer three reveals a dicotomic grouping between the animals from both states in the Yucatan Peninsula showing a divergent value of 1.983 nucleotides per hundred. Animals from Quintana Roo show a grouping with primer six; an additional grouping was observed with animals from Campeche. Nucleotidic variation between both groups was 2.118 nucleotides per hundred. The nucleotidic variation for the two primers within the groups from both states, showed fluctuating values from 0.46 to 1.68 nucleotides per hundred, which indicates that nucleotidic variation between the two groups of animals is around two nucleotides per hundred and, within the groups, less than 1.7 nucleotides per hundred. 相似文献
7.
We describe a new statistical method for measuring bias in the codon usage table of a gene. The test is based on the multinomial and Poisson distributions. The method is used to scan DNA sequences and measure the strength of codon preference. For E. Coli we show that the strength of codon preference is related to levels of gene expression. The method can also be used to compare base triplet frequencies with those expected from the base composition. This second type of codon bias test is useful for distinguishing coding from non-coding regions. 相似文献
8.
Strength of translation initiation signal sequence of mRNA as studied by quantification method: effect of nucleotide substitutions upon translation efficiency in rat preproinsulin mRNA. 总被引:2,自引:1,他引:1
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Concerning the translation initiation signals in vertebrate mRNAs, both the ATG initiation codon and the sequences flanking the initiation codon are required to direct the position of initiation. A consensus sequence for the signal, (GCC)GCC(A or G)CCATGG, has been proposed, but actual initiation sequences differ from it to a greater or lesser degree. In the present report, the translation initiation signal sequences of rat preproinsulin and its mutant mRNAs were analyzed using a quantification method proposed previously. In this method, each 16 nt sequence in the mRNA was characterized by its sample score, which shows strength of the signal. So far, Kozak has constructed a number of preproinsulin mutant mRNAs in which nucleotides flanking the ATG codon are systematically varied, and measured the translation initiation efficiency in terms of the proinsulin product. Her experimental results were well understood on the basis of the strength of the translation initiation signal sequence. 相似文献
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V. A. Efimov I. Y. Dubey O. G. Chakhmakhcheva 《Nucleosides, nucleotides & nucleic acids》2013,32(3):473-477
Abstract The mechanism of the internucleotide condensation and side-reactions in H-phosphonate approach has been investigated. The modification of this method allowed to minimize side-reactions during the preactivation of the nucleotidic component has been developed. 相似文献
10.
This study describes a method for the backtranslation of an aminoacidic sequence, an extremely useful tool for various experimental approaches. It involves two computer programs CLUSTER and BACKTR written in Fortran 77 running on a VAX/VMS computer. CLUSTER generates a reliable codon usage table through a cluster analysis, based on a chi 2-like distance between the sequences. BACKTR produces backtranslated sequences according to different options when use is made of the codon usage table obtained in addition to selecting the least ambiguous potential oligonucleotide probes within an aminoacidic sequence. The method was tested by applying it to 158 yeast genes. 相似文献
11.
We have developed a robust and sensitive method, called RNA-ID, to screen for cis-regulatory sequences in RNA using fluorescence-activated cell sorting (FACS) of yeast cells bearing a reporter in which expression of both superfolder green fluorescent protein (GFP) and yeast codon-optimized mCherry red fluorescent protein (RFP) is driven by the bidirectional GAL1,10 promoter. This method recapitulates previously reported progressive inhibition of translation mediated by increasing numbers of CGA codon pairs, and restoration of expression by introduction of a tRNA with an anticodon that base pairs exactly with the CGA codon. This method also reproduces effects of paromomycin and context on stop codon read-through. Five key features of this method contribute to its effectiveness as a selection for regulatory sequences: The system exhibits greater than a 250-fold dynamic range, a quantitative and dose-dependent response to known inhibitory sequences, exquisite resolution that allows nearly complete physical separation of distinct populations, and a reproducible signal between different cells transformed with the identical reporter, all of which are coupled with simple methods involving ligation-independent cloning, to create large libraries. Moreover, we provide evidence that there are sequences within a 9-nt library that cause reduced GFP fluorescence, suggesting that there are novel cis-regulatory sequences to be found even in this short sequence space. This method is widely applicable to the study of both RNA-mediated and codon-mediated effects on expression. 相似文献
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Kazuyuki Takai 《Nucleosides, nucleotides & nucleic acids》2016,35(5):223-232
Gene synthesis is getting more important with the growing availability of low-cost commercial services. The coding sequences are often “optimized” as for the relative synonymous codon usage (RSCU) before synthesis, which is generally included in the commercial services. However, the codon optimization processes are different among different providers and are often hidden from the users. Here, the d'Hondt method, which is widely adopted as a method for determining the number of seats for each party in proportional-representation public elections, is applied to RSCU fitting. This allowed me to make a set of electronic spreadsheets for manual design of protein coding sequences for expression in Escherichia coli, with which users can see the process of codon optimization and can manually edit the codons after the automatic optimization. The spreadsheets may also be useful for molecular biology education 相似文献
13.
不具有3-碱基周期性的编码序列初探 总被引:4,自引:0,他引:4
对120个较短编码序列(<1 200 bp)的Fourier频谱进行分析表明,3-碱基周期性在短编码序列中并不是绝对存在的.统计分析提示,编码序列有无3-碱基周期性与序列的碱基组成和分布、所编码蛋白质氨基酸的选用和顺序以及同义密码子的使用都有一定的关系.一般地,非周期-3序列中A+U含量高于G+C含量,周期-3序列的情况则相反;非周期-3序列中碱基在密码子三个位点上的分布比周期-3序列中的分布均匀;非周期-3序列密码子和氨基酸的使用偏向没有周期-3序列的大.在利用Fourier分析方法预测DNA序列中的基因和外显子时,应充分考虑到这些现象. 相似文献
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赋予氨基酸编码方法下除终止子之外的密码子突变为终止子时每一位发生的变化权值,利用矩阵来表示所有的突变方式和难易程度,综合亲水性与疏水性理化性质,提出亚氨基酸编码方法,并给出该编码方法下同义密码子的相对使用度fSubtypesRelativeSynonymousCodonUsage,SRSCU).然后选取15条H5N1序列,使用MEGA4.0分析它们的同源性,并分别在氨基酸编码、拟氨基酸编码、亚氨基酸编码这三种环境下研究所选序列使用密码子的偏好性,对比结果验证,亚氨基酸编码方法具有相应的优越性. 相似文献
15.
Methylome profiling of cancer cells by amplification of inter-methylated sites (AIMS) 总被引:5,自引:1,他引:4
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Alterations of the DNA methylation pattern have been related to generalized chromosomal disruption and inactivation of multiple tumor suppressor genes in neoplasia. To screen for tumor-specific alterations and to make a global assessment of methylation status in cancer cells, we have modified the methylated CpG island amplification method to generate easily readable fingerprints representing the cell’s DNA methylation profile. The method is based on the differential cleavage of isoschizomers with distinct methylation sensitivity. Specific adaptors are ligated to the methylated ends of the digested genomic DNA. The ligated sequences are amplified by PCR using adaptor- specific primers extended at the 3′ end with two to four arbitrarily chosen nucleotidic residues to reduce the complexity of the product. Fingerprints consist of multiple anonymous bands, representing DNA sequences flanked by two methylated sites, which can be isolated and individually characterized. Hybridization of the whole product to metaphase chromosomes revealed that most bands originate from the isochore H3, which identifies the regions of the genome with the highest content of CpG islands and genes. Comparison of the fingerprints obtained from normal colon mucosa, colorectal carcinomas and cell lines revealed tumor-specific alterations that are putative recurrent markers of the disease and include tumor-specific hypo- and hypermethylations. 相似文献
16.
Mehmood T Bohlin J Bråthen Kristoffersen A Sæbø S Warringer J Snipen L 《BMC bioinformatics》2012,13(1):97
ABSTRACT: BACKGROUND: Gene finding is a complicated procedure that encapsulates algorithms for coding sequence modeling, identification of promoter regions, issues concerning overlapping genes and more. In the present study we focus on coding sequence modeling algorithms; that is, algorithms for identification and prediction of the actual coding sequences from genomic DNA. In this respect, we promote a novel multivariate method known as Canonical Powered Partial Least Squares (CPPLS) as an alternative to the commonly used Interpolated Markov model (IMM). Comparisons between the methods were performed on DNA, codon and protein sequences with highly conserved genes taken from several species with different genomic properties. RESULTS: The multivariate CPPLS approach classified coding sequence substantially better than the commonly used IMM on the same set of sequences. We also found that the use of CPPLS with codon representation gave significantly better classification results than both IMM with protein (p < 0.001) and with DNA (p < 0.001). Further, although the mean performance was similar, the variation of CPPLS performance on codon representation was significantly smaller than for IMM (p < 0.001). CONCLUSIONS: The performance of coding sequence modeling can be substantially improved by using an algorithm based on the multivariate CPPLS method applied to codon or DNA frequencies. 相似文献
17.
Despite the establishment of design principles to optimize codon choice for heterologous expression vector design, the relationship between codon sequence and final protein yield remains poorly understood. In this work, we present a computational framework for the identification of a set of mutant codon sequences for optimized heterologous protein production, which uses a codon-sequence mechanistic model of protein synthesis. Through a sensitivity analysis on the optimal steady state configuration of protein synthesis we are able to identify the set of codons, that are the most rate limiting with respect to steady state protein synthesis rate, and we replace them with synonymous codons recognized by charged tRNAs more efficient for translation, so that the resulting codon-elongation rate is higher. Repeating this procedure, we iteratively optimize the codon sequence for higher protein synthesis rate taking into account multiple constraints of various types. We determine a small set of optimized synonymous codon sequences that are very close to each other in sequence space, but they have an impact on properties such as ribosomal utilization or secondary structure. This limited number of sequences can then be offered for further experimental study. Overall, the proposed method is very valuable in understanding the effects of the different properties of mRNA sequences on the final protein yield in heterologous protein production and it can find applications in synthetic biology and biotechnology. 相似文献
18.
Bu SL 《General physiology and biophysics》2012,31(2):221-224
Here, we study the topological properties of genetic sequence by viewing the entire sequence as a whole. First, a systematic way of coding the gene-combinations in a genetic sequence is developed. Next, we apply the coding method to real genetic sequences, and find a scale-free power-law distribution for some particular kinds of gene-combinations. Furthermore, we also present a model to reproduce the observed scale-free feature, which is based on three generic mechanisms: 1) Growth mechanism - genetic sequences expands continuously by the addition of new codon; 2) Preferential replication mechanism - the newly added codon at every time step is a replication of one of existed condons, and the probability that a codon is replicated is proportional to its emerging times in existed sequence; and 3) Mutation mechanism - following (2) the newly added codon has a small probability to mutate. To our knowledge, no report has been published to study the genetic sequences in such way. 相似文献
19.
Estimating synonymous and nonsynonymous substitution rates under realistic evolutionary models 总被引:22,自引:0,他引:22
Approximate methods for estimating the numbers of synonymous and nonsynonymous substitutions between two DNA sequences involve three steps: counting of synonymous and nonsynonymous sites in the two sequences, counting of synonymous and nonsynonymous differences between the two sequences, and correcting for multiple substitutions at the same site. We examine complexities involved in those steps and propose a new approximate method that takes into account two major features of DNA sequence evolution: transition/transversion rate bias and base/codon frequency bias. We compare the new method with maximum likelihood, as well as several other approximate methods, by examining infinitely long sequences, performing computer simulations, and analyzing a real data set. The results suggest that when there are transition/transversion rate biases and base/codon frequency biases, previously described approximate methods for estimating the nonsynonymous/synonymous rate ratio may involve serious biases, and the bias can be both positive and negative. The new method is, in general, superior to earlier approximate methods and may be useful for analyzing large data sets, although maximum likelihood appears to always be the method of choice. 相似文献
20.
The mosquito Aedes aegypti is the primary vector of dengue virus (DENV) infection in most of the subtropical and tropical countries. Besides DENV, yellow fever virus (YFV) is also transmitted by A. aegypti. Susceptibility of A. aegypti to West Nile virus (WNV) has also been confirmed. Although studies have indicated correlation of codon bias between flaviviridae and their animal/insect hosts, it is not clear if codon sequences have any relation to susceptibility of A. aegypti to DENV, YFV and WNV. In the current study, usages of codon context sequences (codon pairs for neighboring amino acids) of the vector (A. aegypti) genome as well as the flaviviral genomes are investigated. We used bioinformatics methods to quantify codon context bias in a genome-wide manner of A. aegypti as well as DENV, WNV and YFV sequences. Mutual information statistics was applied to perform bicluster analysis of codon context bias between vector and flaviviral sequences. Functional relevance of the bicluster pattern was inferred from published microarray data. Our study shows that codon context bias of DENV, WNV and YFV sequences varies in a bicluster manner with that of specific sets of genes of A. aegypti. Many of these mosquito genes are known to be differentially expressed in response to flaviviral infection suggesting that codon context sequences of A. aegypti and the flaviviruses may play a role in the susceptible interaction between flaviviruses and this mosquito. The bias in usages of codon context sequences likely has a functional association with susceptibility of A. aegypti to flaviviral infection. The results from this study will allow us to conduct hypothesis-driven tests to examine the role of codon context bias in evolution of vector–virus interactions at the molecular level. 相似文献