Lampe M, Binnie C, Schmidt R, Losick R
Department of Cellular and Developmental Biology, Harvard University, Cambridge, MA 02138.
Gene. 1988 Jul 15;67(1):13-9. doi: 10.1016/0378-1119(88)90003-0.
Core RNA polymerase and several forms of RNA polymerase holoenzyme from Bacillus subtilis are found in association with a 21,500-kDa polypeptide called delta (delta). We have cloned the structural gene (rpoE) for delta by using a hybridization probe a synthetic oligodeoxynucleotide that was designed on the basis of a partial NH2-terminal amino acid (aa) sequence of purified delta protein. The rpoE gene was found to encode a 173-aa polypeptide of a predicted Mr of 20,400. Genetic and physical mapping experiments placed rpoE at 325 degrees on the B. subtilis chromosome and established the gene order rpoE-ctrA-spo0F. The delta subunit is known to enhance the specificity of transcription in vitro by bacterial and phage SP01-modified forms of polymerase, but replacement of rpoE by an in vitro-constructed deletion-mutated gene was found not to impair viability, sporulation, or the growth of phage SP01.
枯草芽孢杆菌的核心RNA聚合酶和几种形式的RNA聚合酶全酶与一种名为δ（delta）的21,500 kDa多肽相关联。我们通过使用杂交探针——一种基于纯化的δ蛋白的部分NH2末端氨基酸（aa）序列设计的合成寡脱氧核苷酸，克隆了δ的结构基因（rpoE）。发现rpoE基因编码一种预测分子量为20,400的173个氨基酸的多肽。遗传和物理图谱实验将rpoE定位在枯草芽孢杆菌染色体的325度处，并确定了基因顺序为rpoE-ctrA-spo0F。已知δ亚基可增强细菌和噬菌体SP01修饰形式的聚合酶在体外的转录特异性，但发现用体外构建的缺失突变基因取代rpoE不会损害噬菌体SP01的活力、孢子形成或生长。
