Gene encoding the 37,000-dalton minor sigma factor of Bacillus subtilis RNA polymerase: isolation, nucleotide sequence, chromosomal locus, and cryptic function.
Gene encoding the 37,000-dalton minor sigma factor of Bacillus subtilis RNA polymerase: isolation, nucleotide sequence, chromosomal locus, and cryptic function.
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编码枯草芽孢杆菌 RNA 聚合酶 37,000 道尔顿小西格玛因子的基因：分离、核苷酸序列、染色体位点和隐秘功能。
DOI：
10.1128/jb.169.2.771-778.1987
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发表时间：
1987
影响因子：
3.2
通讯作者：
Price,CW
中科院分区：
文献类型：
--
作者：
Duncan,ML;Kalman,SS;Thomas,SM;Price,CW
我们开始对rpoF进行分析，rpoF是枯草芽孢杆菌RNA聚合酶的一种编码37，000-道尔顿小σ因子（σ-37）的基因。使用针对σ-37全酶的抗体来探测λ gt 11表达载体文库，我们分离了表达与lacZ融合的σ-37的COOH末端的一半的901个碱基对的EcoRI片段。我们使用该片段作为杂交探针来分离整个rpoF基因和另外的侧翼序列。克隆基因的身份通过其在大肠杆菌中表达的产物的大小和免疫反应以及DNA测序后其预测产物（264个残基; 30，143道尔顿）与其他σ因子的同源性来确认。DNA序列分析还表明rpoF可能位于一个基因簇中。rpoF的上游是一个开放的阅读框架，它将编码17，992道尔顿的蛋白质;该框架与rpoF编码序列重叠41个碱基对。紧接着rpoF的是一个阅读框，它将编码至少20，000道尔顿的蛋白质;该区域的表达可能与rpoF的表达偶联。通过质粒整合和PBS 1转导，我们发现rpoF的染色体位点与B上的ddl和dal以40度连锁。枯草杆菌地图和几乎没有已知的病变影响生长调节或发展。此外，由基因破坏引起的rpoF无效突变对丰富培养基中的细胞生长或孢子形成没有影响，表明σ-37可能部分控制不直接参与孢子形成过程的调节子。
We began an analysis of rpoF, the gene encoding the cryptic, 37,000-dalton minor sigma factor (sigma-37) of Bacillus subtilis RNA polymerase. Using antibody raised against sigma-37 holoenzyme to probe a lambda gt11 expression vector library, we isolated a 901-base-pair EcoRI fragment that expressed the COOH-terminal half of sigma-37 fused to lacZ. We used this fragment as a hybridization probe to isolate the entire rpoF gene and additional flanking sequences. Identity of the cloned gene was confirmed by the size and immunological reaction of its product expressed in Escherichia coli and, after DNA sequencing, by the homology of its predicted product (264 residues; 30,143 daltons) with other sigma factors. The DNA sequence also suggested that rpoF may lie in a gene cluster. Upstream of rpoF was an open reading frame that would encode a protein of 17,992 daltons; this frame overlapped the rpoF-coding sequence by 41 base pairs. Immediately following rpoF was a reading frame that would encode a protein of at least 20,000 daltons; expression of this region may be translationally coupled to that of rpoF. By plasmid integration and PBS1 transduction, we found the chromosomal locus of rpoF linked to ddl and dal at 40 degrees on the B. subtilis map and near no known lesions affecting growth regulation or development. Further, an rpoF null mutation resulting from gene disruption had no effect on cell growth or sporulation in rich medium, suggesting that sigma-37 may partly control a regulon not directly involved in the sporulation process.