Concurrent Growth Rate and Transcript Analyses Reveal Essential Gene Stringency in Escherichia coli
Concurrent Growth Rate and Transcript Analyses Reveal Essential Gene Stringency in Escherichia coli
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DOI：
10.1371/journal.pone.0006061
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发表时间：
2009-06-26
期刊：
影响因子：
3.7
通讯作者：
Good, Liam
中科院分区：
文献类型：
--
作者：
Goh, Shan;Boberek, Jaroslaw M.;Good, Liam
背景：细菌生长所必需的基因具有特别的科学意义。许多假定的必需基因已被确定或预测在几个物种中，然而，鲜为人知的是基因表达的要求严格性，这可能是细菌生理学的一个重要方面，并可能在药物靶点development.Methodology/Principal调查结果的决定因素：工作的前提下，必需基因不同的绝对要求的增长，我们描述了沉默假定的必需基因在大肠杆菌。用反义肽核酸（PNA）和表达的反义RNA对大肠杆菌的生长速率和转录水平进行滴定。mRNA下降和生长速率下降之间的关系反映了必需基因的必要性或严格性的程度，其在此由生长速率降低50%的最小转录水平（MTL 50）定义。当应用于四个生长必需基因时，两种RNA沉默方法导致MTL 50值，其揭示acpP是所检查的四个基因中最严格需要的，ftsZ是下一个最严格需要的。已建立的抗菌目标murA和fabI不太严格required.Conclusions：RNA沉默可以揭示严格的要求基因表达相对于生长。该方法可用于验证现有的必需基因和定量药物靶向需求。
Background: Genes essential for bacterial growth are of particular scientific interest. Many putative essential genes have been identified or predicted in several species, however, little is known about gene expression requirement stringency, which may be an important aspect of bacterial physiology and likely a determining factor in drug target development.Methodology/Principal Findings: Working from the premise that essential genes differ in absolute requirement for growth, we describe silencing of putative essential genes in E. coli to obtain a titration of declining growth rates and transcript levels by using antisense peptide nucleic acids (PNA) and expressed antisense RNA. The relationship between mRNA decline and growth rate decline reflects the degree of essentiality, or stringency, of an essential gene, which is here defined by the minimum transcript level for a 50% reduction in growth rate (MTL50). When applied to four growth essential genes, both RNA silencing methods resulted in MTL50 values that reveal acpP as the most stringently required of the four genes examined, with ftsZ the next most stringently required. The established antibacterial targets murA and fabI were less stringently required.Conclusions: RNA silencing can reveal stringent requirements for gene expression with respect to growth. This method may be used to validate existing essential genes and to quantify drug target requirement.