Hui F M, Morrison D A
Department of Biological Sciences, University of Illinois, Chicago 60680.
J Bacteriol. 1991 Jan;173(1):372-81. doi: 10.1128/jb.173.1.372-381.1991.
The complete nucleotide sequence of comA, a gene required for induction of competence for genetic transformation in Streptococcus pneumoniae, was determined by using plasmid DNA templates and synthetic oligonucleotide primers. The sequence contained a single large open reading frame, ORF1, of 2,151 bp. ORF1 was included within the comAB locus previously mapped genetically and accounted for 50% of its extent. The predicted molecular weight of the largest polypeptide encoded within ORF1, 80,290, coincided with that measured previously (77,000) for the product of in vitro transcription-translation of the cloned comA locus. A Shine-Dalgarno sequence (AAAGGAG, delta G = -14 kcal) lay immediately upstream of ORF1. A sequence (TTtAat-17 bp-TAaAAT) similar to the Escherichia coli sigma 70 promoter consensus was located 410 bp upstream of ORF1. The deduced protein sequence of ComA showed a very strong similarity to the E. coli hemolysin secretion protein, HlyB, and strong similarities to other members of the family of ATP-dependent transport proteins, including the mammalian multidrug resistance P-glycoprotein. These similarities suggest that ComA functions in the transport of some molecule, possibly pneumococcal competence factor itself.
通过使用质粒DNA模板和合成寡核苷酸引物,确定了肺炎链球菌中遗传转化感受态诱导所需基因comA的完整核苷酸序列。该序列包含一个2151 bp的单一大型开放阅读框ORF1。ORF1包含在先前通过遗传学方法定位的comAB基因座内,占其长度的50%。ORF1内编码的最大多肽的预测分子量为80290,与先前测量的克隆comA基因座体外转录-翻译产物的分子量(77000)一致。一个Shine-Dalgarno序列(AAAGGAG,ΔG = -14 kcal)位于ORF1的紧邻上游。一个与大肠杆菌σ70启动子共有序列相似的序列(TTtAat - 17 bp - TAaAAT)位于ORF1上游410 bp处。推导的ComA蛋白序列与大肠杆菌溶血素分泌蛋白HlyB有非常强的相似性,与ATP依赖性转运蛋白家族的其他成员也有很强的相似性,包括哺乳动物多药耐药P-糖蛋白。这些相似性表明ComA在某种分子的转运中起作用,可能是肺炎链球菌感受态因子本身。