分枝杆菌中条件性基因敲低突变体的构建。

Construction of conditional knockdown mutants in mycobacteria.

作者信息

Schnappinger Dirk, O'Brien Kathryn M, Ehrt Sabine

机构信息

Department of Microbiology and Immunology, Weill Cornell Medical College, 1300 York Avenue, New York, NY, 10065, USA,

出版信息

Methods Mol Biol. 2015;1285:151-75. doi: 10.1007/978-1-4939-2450-9_9.

DOI:10.1007/978-1-4939-2450-9_9

PMID:25779315

Abstract

By definition, essential genes are fundamental to bacterial growth, yet the functions of many such genes remain unknown. Essential genes furthermore are central to the activity of most antibacterial drugs and among the most attractive targets for the development of new therapeutics. This chapter describes how synthetic genetic switches that utilize transcriptional repression, controlled proteolysis, or both to silence gene activity can be applied to construct and characterize conditional knockdown (cKD) mutants for essential genes in Mycobacterium smegmatis and Mycobacterium tuberculosis.

摘要

根据定义，必需基因是细菌生长所必需的，但许多此类基因的功能仍不为人知。此外，必需基因是大多数抗菌药物活性的核心，也是开发新疗法最具吸引力的靶点之一。本章描述了如何应用利用转录抑制、可控蛋白水解或两者兼用来沉默基因活性的合成基因开关，构建和表征耻垢分枝杆菌和结核分枝杆菌中必需基因的条件性敲低（cKD）突变体。