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肺炎链球菌密码子偏倚基因、微阵列表达值与生理特征之间的关系

Relationship between codon biased genes, microarray expression values and physiological characteristics of Streptococcus pneumoniae.

作者信息

Martín-Galiano Antonio J, Wells Jerry M, de la Campa Adela G

机构信息

Unidad de Genética Bacteriana (CSIC), Centro Nacional de Microbiología, Instituto de Salud Carlos III, 28220, Majadahonda, Madrid, Spain.

Bacterial Infection and Immunity Group, Institute of Food Research, Norwich Research Park, Norwich NR4 7UA, UK.

出版信息

Microbiology (Reading). 2004 Jul;150(Pt 7):2313-2325. doi: 10.1099/mic.0.27097-0.

Abstract

A codon-profile strategy was used to predict gene expression levels in Streptococcus pneumoniae. Predicted highly expressed (PHE) genes included those encoding glycolytic and fermentative enzymes, sugar-conversion systems and carbohydrate-transporters. Additionally, some genes required for infection that are involved in oxidative metabolism and hydrogen peroxide production were PHE. Low expression values were predicted for genes encoding specific regulatory proteins like two-component systems and competence genes. Correspondence analysis localized 484 ORFs which shared a distinctive codon profile in the right horn. These genes had a mean G+C content (33.4 %) that was lower than the bulk of the genome coding sequences (39.7 %), suggesting that many of them were acquired by horizontal transfer. Half of these genes (242) were pseudogenes, ORFs shorter than 80 codons or without assigned function. The remaining genes included several virulence factors, such as capsular genes, iga, lytB, nanB, pspA, choline-binding proteins, and functions related to DNA acquisition, such as restriction-modification systems and comDE. In order to compare predicted translation rate with the relative amounts of mRNA for each gene, the codon adaptation index (CAI) values were compared with microarray fluorescence intensity values following hybridization of labelled RNA from laboratory-grown cultures. High mRNA amounts were observed in 32.5 % of PHE genes and in 64 % of the 25 genes with the highest CAI values. However, high relative amounts of RNA were also detected in 10.4 % of non-PHE genes, such as those encoding fatty acid metabolism enzymes and proteases, suggesting that their expression might also be regulated at the level of transcription or mRNA stability under the conditions tested. The effects of codon bias and mRNA amount on different gene groups in S. pneumoniae are discussed.

摘要

采用密码子谱策略预测肺炎链球菌中的基因表达水平。预测的高表达(PHE)基因包括那些编码糖酵解和发酵酶、糖转化系统及碳水化合物转运蛋白的基因。此外,一些参与氧化代谢和过氧化氢产生的感染所需基因也是高表达基因。预测编码特定调控蛋白(如双组分系统和感受态基因)的基因表达值较低。对应分析定位了484个在右角共享独特密码子谱的开放阅读框(ORF)。这些基因的平均G+C含量(33.4%)低于基因组编码序列的大部分(39.7%),这表明其中许多基因是通过水平转移获得的。这些基因中有一半(242个)是假基因,即开放阅读框短于80个密码子或无指定功能。其余基因包括几种毒力因子,如荚膜基因、iga、lytB、nanB、pspA、胆碱结合蛋白,以及与DNA获取相关的功能,如限制修饰系统和comDE。为了比较预测的翻译速率与每个基因的mRNA相对量,将密码子适应指数(CAI)值与来自实验室培养物的标记RNA杂交后的微阵列荧光强度值进行比较。在32.5%的高表达基因以及25个CAI值最高的基因中的64%中观察到高mRNA量。然而,在10.4%的非高表达基因中也检测到高相对量的RNA,如那些编码脂肪酸代谢酶和蛋白酶的基因,这表明在测试条件下它们的表达也可能在转录或mRNA稳定性水平受到调控。本文讨论了密码子偏好和mRNA量对肺炎链球菌不同基因组的影响。

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