ISOLATION OF PROKARYOTIC RNA AND DETECTION OF SPECIFIC MESSENGER-RNA WITH BIOTINYLATED PROBES

被引：143

作者：

OELMULLER, U ^{[1
]}

KRUGER, N ^{[1
]}

STEINBUCHEL, A ^{[1
]}

FRIEDRICH, CG ^{[1
]}

机构：

[1] TECH FACHHSCH BERLIN,BERLIN,GERMANY

来源：

JOURNAL OF MICROBIOLOGICAL METHODS | 1990年 / 11卷 / 01期

关键词：

Alcaligenes eutrophus; Biotinylated probe; Hydrogenase; mRNA; Poly(β-hydroxybutyrate);

D O I：

10.1016/0167-7012(90)90050-G

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A rapid method is presented for the isolation of total RNA of prokaryotic cells and the identification of specific mRNA with high sensitivity using a modified method for the detection of bioytinylated DNA. Total RNA was isolated by a procedure based on the principles of nucleic acid isolation. A significant reduction in mRNA degradation was achieved by (i) a drastic reduction in time for cell separation and (ii) employing a phenol-choloroform-SDS mixture of RNA extraction. RNA was transferred to positively charged nylon membranes and for the first time biotinylated probes were applied for mRNA identification. Genes of Alcaligenes eutrophus of soluble hydrogenase (hoxS) and of the ezymes involved in poly(β-hydroybutyrate) formation were used for the establishment of the method. © 1990.

引用

页码：73 / 81

页数：9

共 25 条

[1] MECHANISMS OF MESSENGER-RNA DECAY IN BACTERIA - A PERSPECTIVE [J].