Suh J W, Boylan S A, Price C W
J Bacteriol. 1986 Oct;168(1):65-71. doi: 10.1128/jb.168.1.65-71.1986.
We isolated the gene encoding the alpha subunit of Bacillus subtilis RNA polymerase from a lambda gt11 expression vector library by using anti-alpha antibody as a probe. Four unique clones were isolated, one carrying a lacZ-alpha gene fusion and three carrying the entire alpha coding region together with additional sequences upstream. The identity of the cloned alpha gene was confirmed by the size and immunological reactivity of its product expressed in Escherichia coli. Further, a partial DNA sequence found the predicted NH2 terminus of alpha homologous with E. coli alpha. By plasmid integration and PBS1 transduction, we mapped alpha near rpsE and within the major ribosomal protein gene cluster on the B. subtilis chromosome. Additional DNA sequencing identified rpsM (encoding S13) and rpsK (encoding S11) upstream of alpha, followed by a 180-base-pair intercistronic region that may contain two alpha promoters. Although the organization of the alpha region resembles that of the alpha operon of E. coli, the putative promoters and absence of rpsD (encoding S4) immediately preceding the B. subtilis alpha gene suggest a different regulation.
我们以抗α抗体为探针,从λgt11表达载体文库中分离出编码枯草芽孢杆菌RNA聚合酶α亚基的基因。分离出了四个独特的克隆,一个携带lacZ-α基因融合体,三个携带完整的α编码区以及上游的其他序列。通过在大肠杆菌中表达的产物的大小和免疫反应性,证实了克隆的α基因的身份。此外,部分DNA序列发现α的预测NH2末端与大肠杆菌α同源。通过质粒整合和PBS1转导,我们将α定位在枯草芽孢杆菌染色体上rpsE附近且在主要核糖体蛋白基因簇内。进一步的DNA测序确定了α上游的rpsM(编码S13)和rpsK(编码S11),随后是一个180个碱基对的基因间区域,该区域可能包含两个α启动子。尽管α区域的组织类似于大肠杆菌的α操纵子,但推测的启动子以及枯草芽孢杆菌α基因之前不存在rpsD(编码S4)表明其调控方式不同。