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枯草芽孢杆菌孢子形成早期表达基因的鉴定。

Identification of Bacillus subtilis genes expressed early during sporulation.

作者信息

Mathiopoulos C, Sonenshein A L

机构信息

Department of Molecular Biology and Microbiology, Tufts University Health Sciences Campus, Boston, Massachusetts 02111.

出版信息

Mol Microbiol. 1989 Aug;3(8):1071-81. doi: 10.1111/j.1365-2958.1989.tb00257.x.

Abstract

Labelled cDNA transcribed in vitro from early-sporulation RNA was enriched for sporulation-specific sequences by subtractive hybridization to an excess of vegetative RNA and used to probe libraries of Bacillus subtilis chromosomal DNA. From the initial collection of clones that coded for RNAs transcribed preferentially during sporulation, several were subcloned and studied in more detail. It was found that two clones contained sequences (dciA and dciB) that had an undetectable level of transcription during vegetative growth but had transcripts that started to appear no later than eight minutes after induction of sporulation. A third DNA segment (dciC) was expressed at a low level in vegetative cells and increased within four minutes after induction of sporulation. The effects of spoO mutations, i.e. mutations that prevent cells from reaching stage I of the sporulation process, were tested. Induction of the dciA and dciB transcripts was significantly reduced in strains carrying mutations in the spoOA and spoOH genes but not in a spoOB mutant strain. In addition, a product of the abrB locus, a locus in which mutations are known to partially overcome the pleiotropic effect of spoOA and spoOB mutations, seemed to be required for dciA and dciB expression.

摘要

从早期芽孢形成RNA体外转录的标记cDNA,通过与过量的营养期RNA进行消减杂交,富集芽孢形成特异性序列,并用于探测枯草芽孢杆菌染色体DNA文库。从最初编码在芽孢形成过程中优先转录的RNA的克隆集合中,挑选了几个进行亚克隆并进行更详细的研究。结果发现,两个克隆包含的序列(dciA和dciB)在营养生长期间转录水平检测不到,但转录本在芽孢形成诱导后不迟于8分钟开始出现。第三个DNA片段(dciC)在营养细胞中低水平表达,并在芽孢形成诱导后4分钟内增加。测试了spoO突变的影响,即阻止细胞进入芽孢形成过程第一阶段的突变。在spoOA和spoOH基因发生突变的菌株中,dciA和dciB转录本的诱导显著降低,但在spoOB突变菌株中没有降低。此外,abrB位点的一个产物,已知该位点的突变可部分克服spoOA和spoOB突变的多效性效应,似乎是dciA和dciB表达所必需的。

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