New method for RNA isolation from actinomycetes: Application to the transcriptional analysis of the alcohol oxidoreductase gene thcE in Rhodococcus and Mycobacterium

被引：8

作者：

Nagy, I ^{[1
]}

Schoofs, G ^{[1
]}

DeSchrijver, A ^{[1
]}

Vanderleyden, J ^{[1
]}

DeMot, R ^{[1
]}

机构：

[1] CATHOLIC UNIV LEUVEN,FA JANSSENS LAB GENET,B-3001 HEVERLEE,BELGIUM

来源：

LETTERS IN APPLIED MICROBIOLOGY | 1997年 / 25卷 / 01期

关键词：

D O I：

10.1046/j.1472-765X.1997.00179.x

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

A new protocol for the isolation of RNA from Rhodococcus and other actinomycetes such as Mycobacterium and Amycolatopsis was developed. The method is based on rapid lysis of cells in a high-speed reciprocal shaker using small abrasive particles followed by spin column purification of the lysate. This quick procedure yields RNA preparations suitable for functional studies. This was shown for the thcE gene of R. erythropolis N186/21, which encodes a N,N'-dimethyl-4-nitrosoaniline-dependent alcohol oxidoreductase. The thcE transcript was detected by Northern hybridization in ii. erythropolis, R. fascians, Mycobacterium chlorophenolicum and Mycobacterium smegmatis. The transcriptional start point of the gene was determined by primer extension of the R. erythropolis mRNA.

引用

页码：75 / 79

页数：5