幽门螺杆菌同源突变株的构建。

Generation of Isogenic Mutant Strains of Helicobacter pylori.

机构信息

Material Measurement Laboratory, National Institute of Standards and Technology, Gaithersburg, MD, USA.

Department of Microbiology and Immunology, Uniformed Services University of the Health Sciences, Bethesda, MD, USA.

出版信息

Methods Mol Biol. 2021;2283:107-122. doi: 10.1007/978-1-0716-1302-3_12.

DOI:10.1007/978-1-0716-1302-3_12

PMID:33765314

Abstract

One important tool in the study of gene function is the construction of mutant strains. Specifically, the construction of isogenic mutant strains imparts researchers with the ability to compare a wild-type strain to a strain that is genetically identical with the exception of the gene of interest. For a bacterial species such as Helicobacter pylori , which is notorious for the genetic heterogeneity seen across isolates, comparisons between isogenic and parental strains control for the genetic variation seen between distinct isolates. This chapter details the construction of a clean gene deletion in which the entire coding region is replaced with a selectable marker. The approach detailed herein allows for the thorough investigation of gene function in the absence of confounding genetic variability.

摘要

在研究基因功能时，一个重要的工具是构建突变株。具体来说，构建同基因的突变株使研究人员能够将野生型菌株与除了感兴趣基因之外在遗传上完全相同的菌株进行比较。对于像幽门螺杆菌这样的细菌物种，由于在分离株中存在遗传异质性而臭名昭著，因此同基因和亲本菌株之间的比较可以控制不同分离株之间的遗传变异。本章详细介绍了构建一个清洁基因缺失的方法，其中整个编码区被一个可选择的标记所取代。本文详细介绍的方法允许在没有混杂遗传变异性的情况下对基因功能进行深入研究。