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质粒R100的traM和finP基因的克隆、定位及测序

Cloning, mapping, and sequencing of plasmid R100 traM and finP genes.

作者信息

Fee B E, Dempsey W B

出版信息

J Bacteriol. 1986 Jul;167(1):336-45. doi: 10.1128/jb.167.1.336-345.1986.

Abstract

The fertility control gene finP, the transfer gene traM, and the transfer origin, oriT, of plasmid R100 were isolated on a single 1.2-kilobase EcoRV fragment and were then subcloned as HaeIII fragments. The sequence of the 754-base-pair finP-containing fragment is reported here. In addition to the finP gene, the sequence includes all but two bases of the R100 traM open reading frame and apparently all of the leader mRNA sequence and amino end of the traJ gene of R100. The sequence contains two open reading frames which encode small proteins on the opposite strand from the traM and traJ genes. It also shows two sets of inverted repeats that have the characteristics of transcription terminators. One set is positioned as if it was the traM terminator, and the other set, which is downstream from the first, sits in the middle of the leader mRNA sequence for traJ. On the bottom strand, this inverted repeat has the structure of a rho-independent terminator. Other less-stable inverted repeats overlap this second terminator in the same way as is seen in attenuation sequences, and the two separate small open reading frames on the bottom strand also totally overlap the stem of the rho-independent terminator, suggesting that their translation would cause shifting of termination to the bottom strand homolog of the putative traM terminator. The finP gene product was not identified, but the gene was mapped to the sequence which contains the traJ gene. It either overlaps traJ or is antisense to it.

摘要

质粒R100的生育控制基因finP、转移基因traM和转移起始位点oriT被分离在一个1.2千碱基的EcoRV片段上,然后作为HaeIII片段进行亚克隆。本文报道了包含finP的754碱基对片段的序列。除了finP基因外,该序列还包括R100 traM开放阅读框中除两个碱基外的所有碱基,以及R100 traJ基因的明显所有前导mRNA序列和氨基末端。该序列包含两个开放阅读框,它们在与traM和traJ基因相反的链上编码小蛋白质。它还显示出两组具有转录终止子特征的反向重复序列。一组的位置好像是traM终止子,另一组在第一组的下游,位于traJ前导mRNA序列的中间。在底部链上,这个反向重复序列具有不依赖ρ因子的终止子结构。其他不太稳定的反向重复序列以与衰减序列中相同的方式与第二个终止子重叠,并且底部链上两个单独的小开放阅读框也完全与不依赖ρ因子的终止子的茎重叠,这表明它们的翻译会导致终止转移到假定的traM终止子底部链的同源物上。finP基因产物未被鉴定,但该基因被定位到包含traJ基因的序列上。它要么与traJ重叠,要么与之反义。

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