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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >Nucleotide sequence of a 7-kb fragment of pACM1 encoding an IncM DNA primase and other putative proteins associated with conjugation.
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Nucleotide sequence of a 7-kb fragment of pACM1 encoding an IncM DNA primase and other putative proteins associated with conjugation.

机译：pACM1的7 kb片段的核苷酸序列，编码IncM DNA primase和其他与结合相关的推定蛋白质。

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摘要

A 7-kb fragment of pACM1 (fragment 9091) containing one or more kor (kill-override) loci was sequenced, and 28 open reading frames (ORFs; >/=50 codons) were identified. The nucleotide sequence has no significant homologs in the GenBank database except for a 1.3-kb region 98.6% identical to the iml (insensitivity to phage PhiM-mediated lysis) determinant fragment of IncM plasmid R446. Deduced amino acid sequences for several ORFs are homologous to those of known proteins, including the Sog DNA primases of IncI1 plasmids R64 and ColIb-P9 and the TraL, TraM, and TraN products of ColIb-P9. Two protein products of the putative primase ORF (ORF 1, 1100 amino acids) were detected by SDS-PAGE. The 158- and 107-kDa proteins were designated PriL and PriS, respectively. PriS is apparently produced by an in-frame reinitiation of the ORF 1 transcript at a second start codon located between a Sau96I site and a PstI site. The motif EGYATA, conserved among primases and associated with primase function, occurs in the first one-third of the deduced amino acid sequence of PriL and is not included in PriS. Partial suppression of the temperature-sensitive dnaG3 mutation in BW86 was demonstrated by recombinants that overexpressed both PriL and PriS, but not by constructs overexpressing only PriS. Therefore, primase function can be assigned to PriL. Fragment 90/91 represents a portion of the IncM tra region, which has not previously been examined in detail. Copyright 2000 Academic Press.
机译：对包含一个或多个kor（杀灭覆盖）位点的pACM1的7kb片段（片段90 91）进行测序，并鉴定出28个开放阅读框（ORF;> / = 50个密码子）。除了与IncM质粒R446的iml决定簇片段的iml（对噬菌体PhiM介导的裂解不敏感）的1.3kb区域98.6％相同外，该核苷酸序列在GenBank数据库中没有明显的同源物。推导的几种ORF的氨基酸序列与已知蛋白的氨基酸序列同源，包括IncI1质粒R64和ColIb-P9的Sog DNA primase和ColIb-P9的TraL，TraM和TraN产物。通过SDS-PAGE检测到推定的primase酶ORF的两种蛋白质产物（ORF 1，1100个氨基酸）。 158-kDa和107-kDa蛋白分别命名为PriL和PriS。显然，PriS是通过在Sau96I位点和PstI位点之间的第二个起始密码子处ORF 1转录本的框内重新初始化产生的。 EGYATA基序在primase中保守并且与primase功能相关，它出现在PriL推导的氨基酸序列的前三分之一中，不包含在PriS中。 BW86中对温度敏感的dnaG3突变的部分抑制作用通过过表达PriL和PriS的重组体得到证实，而没有通过仅过表达PriS的构建体得到证明。因此，可以将Primase功能分配给PriL。片段90/91代表IncM tra区域的一部分，之前未进行过详细检查。版权所有2000学术出版社。

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