Sorokin A, Zumstein E, Azevedo V, Ehrlich S D, Serror P
Laboratoire de Génétique Microbienne, Institut National de la Recherche Agronomique, Jouy en Josas, France.
Mol Microbiol. 1993 Oct;10(2):385-95. doi: 10.1111/j.1365-2958.1993.tb02670.x.
Three different lambda phage clones with overlapping inserts of Bacillus subtilis DNA, which cover the region from spoIIAA to serA, have been isolated. The nucleotide sequence of their inserts, starting after spoVAF and ending at serA, has been determined. A contiguous sequence of 35,354 bp was established, including previously analysed overlapping adjacent regions. Within the newly determined sequence 31 open reading frames (ORFs) with putative ribosome-binding sites have been found. Nine of them correspond to previously sequenced and characterized genes: spo-VAF, lysA, sipS, ribG, ribB, ribA, ribH, ribTD and dacB. Comparison of the amino acid sequences of the products encoded by the other ORFs to known proteins allowed putative functions to be assigned to seven of these ORFs. Among these are the following: (i) the ppiB gene, encoding a cytoplasmic peptidylprolyl isomerase; (ii) two pairs of signal-transducers, one homologous to phoR-phoP of B. subtilis, encoding regulators of phosphatase biosynthesis, and the second to the fecI-fecR of Escherichia coli, which is responsible for the regulation of the citrate-dependent iron (III) transport system; (iii) aroC and serA genes, involved in the biosynthesis of aromatic amino acids and serine, respectively, the function of which has been confirmed by constructing corresponding mutants with disrupted ORFs. The organization of putative operons has been postulated on the basis of the sequences of their transcription terminators, promoters and regulatory elements.
已分离出三个不同的λ噬菌体克隆,其插入的枯草芽孢杆菌DNA片段相互重叠,覆盖了从spoIIAA到serA的区域。已确定了其插入片段从spoVAF之后开始到serA结束的核苷酸序列。建立了一个35354 bp的连续序列,包括先前分析过的重叠相邻区域。在新确定的序列中,发现了31个带有推定核糖体结合位点的开放阅读框(ORF)。其中9个对应于先前测序和表征的基因:spo-VAF、lysA、sipS、ribG、ribB、ribA、ribH、ribTD和dacB。将其他ORF编码产物的氨基酸序列与已知蛋白质进行比较,为其中7个ORF赋予了推定功能。其中包括:(i)ppiB基因,编码一种细胞质肽基脯氨酰异构酶;(ii)两对信号转导器,一对与枯草芽孢杆菌的phoR-phoP同源,编码磷酸酶生物合成的调节因子,另一对与大肠杆菌的fecI-fecR同源,负责柠檬酸依赖性铁(III)转运系统调节;(iii)aroC和serA基因,分别参与芳香族氨基酸和丝氨酸的生物合成,通过构建相应的ORF破坏突变体已证实了它们的功能。已根据推定操纵子的转录终止子、启动子和调节元件序列推测了其组织方式。