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大肠杆菌致育因子F的traB - traC区域内转移基因及基因产物的分析

Analysis of transfer genes and gene products within the traB-traC region of the Escherichia coli fertility factor, F.

作者信息

Moore D, Wu J H, Kathir P, Hamilton C M, Ippen-Ihler K

出版信息

J Bacteriol. 1987 Sep;169(9):3994-4002. doi: 10.1128/jb.169.9.3994-4002.1987.

Abstract

A series of plasmids that carry overlapping segments of F DNA encoding the genes in the traB-traC interval was constructed, and a restriction enzyme map of the region was derived. Plasmids carrying deletions that had been introduced at an HpaI site within this interval were also isolated. The ability of these plasmids to complement transfer of F lac plasmids carrying mutations in traB, traV, and traW, and traC was analyzed. The protein products of the plasmids were labeled in UV-irradiated cells and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. These analyses showed that the product of traV is a polypeptide that migrates with an apparent molecular weight of 21,000. It was not detected when [35S]methionine was used to label plasmid products, but was readily detected in 14C-amino acid labeling experiments. A 21,500-dalton product appeared to stem from the region assigned to traP. A 9,000-dalton product was found to stem from a locus, named traR, that is located between traV and traC. No traW activity could be detected from the region of tra DNA examined. Our data also indicated that traC is located in a more promoter-proximal position than suggested on earlier maps. The plasmids constructed are expected to be useful in studies designed to identify the specific functions of the traB, -P, -V, -R, and -C products.

摘要

构建了一系列携带F DNA重叠片段的质粒,这些片段编码traB - traC区间内的基因,并得到了该区域的限制性酶切图谱。还分离出了携带在此区间内HpaI位点引入缺失的质粒。分析了这些质粒对携带traB、traV、traW和traC突变的F lac质粒转移的互补能力。在紫外线照射的细胞中对质粒的蛋白质产物进行标记,并通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳和放射自显影进行分析。这些分析表明,traV的产物是一种多肽,其表观分子量为21,000,在使用[35S]甲硫氨酸标记质粒产物时未检测到,但在14C - 氨基酸标记实验中很容易检测到。一个21,500道尔顿的产物似乎源于分配给traP的区域。发现一个9,000道尔顿的产物源于一个位于traV和traC之间的名为traR的位点。在所检测的tra DNA区域未检测到traW活性。我们的数据还表明,traC位于比早期图谱所显示的更靠近启动子的位置。所构建的质粒有望用于旨在鉴定traB、 - P、 - V、 - R和 - C产物特定功能的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01ca/213699/88c7d233cec5/jbacter00199-0144-a.jpg

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