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用于枯草芽孢杆菌的gusA转录融合载体的构建及其在孢子形成研究中的应用。

Construction of gusA transcriptional fusion vectors for Bacillus subtilis and their utilization for studies of spore formation.

作者信息

Karow M L, Piggot P J

机构信息

Department of Microbiology and Immunology, Temple University School of Medicine, Philadelphia, PA 19140, USA.

出版信息

Gene. 1995 Sep 22;163(1):69-74. doi: 10.1016/0378-1119(95)00402-r.

Abstract

A series of gusA transcriptional fusion vectors is described for Bacillus subtilis (Bs). The series includes a vector for use with the amyE system of Shimotsu and Henner [Gene 43 (1986) 85-94], an integrative vector and vectors that provide gusA or gusA neo cassettes. The gusA fusions are compatible with lacZ fusion vectors that are widely used with Bs, and gusA and lacZ fusions are expressed at similar levels. beta-Glucuronidase (beta Glu) and beta-galactosidase (beta Gal) do not exhibit any cross-reactivity, there is very little endogenous beta Glu activity in Bs, and there is no indication of mutation to high-level expression. We have use strains containing both gusA and lacZ fusions to compare the times of expression of different genes during sporulation.

摘要

本文描述了一系列用于枯草芽孢杆菌(Bs)的gusA转录融合载体。该系列包括一个与Shimotsu和Henner [《基因》43 (1986) 85 - 94]的amyE系统一起使用的载体、一个整合载体以及提供gusA或gusA neo盒式结构的载体。gusA融合体与广泛用于枯草芽孢杆菌的lacZ融合载体兼容,并且gusA和lacZ融合体以相似水平表达。β-葡萄糖醛酸酶(β-Glu)和β-半乳糖苷酶(β-Gal)没有表现出任何交叉反应性,枯草芽孢杆菌中内源性β-Glu活性非常低,并且没有向高水平表达突变的迹象。我们已经使用了同时含有gusA和lacZ融合体的菌株来比较芽孢形成过程中不同基因的表达时间。

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