Sarandopoulos S, Davies J K
Department of Microbiology, Monash University, Clayton, Victoria, Australia.
Plasmid. 1993 May;29(3):206-21. doi: 10.1006/plas.1993.1023.
Three promoters have been identified on the phenotypically cryptic plasmid of Neisseria gonorrhoeae that can direct transcription in Escherichia coli. These promoters do not seem to function at a detectable level when N. gonorrhoeae is grown in vitro under normal growth conditions. The T7 RNA polymerase expression system has been used to identify the proteins encoded by this plasmid, and a series of subclones were constructed and used to localize the gene encoding each protein. In a search of the sequence databases, we have discovered that the derived amino acid sequence of a 1.2-kb segment of the plasmid shows a high degree of sequence similarity to the Mob proteins encoded by the colicinogenic plasmids ColA, ColE1, and ColK of E. coli. The 1.2-kb segment contains part of a Col mobilization region that has subsequently been inactivated by a small duplication. During the recombination event that formed the cryptic plasmid one of the mobilization genes was fused to a cryptic plasmid open reading frame to form the cppB gene. Despite the manner in which it was formed, we have shown that the cppB gene can be expressed when N. gonorrhoeae is grown under the appropriate environmental conditions.
在淋病奈瑟菌的表型隐匿质粒上已鉴定出三个启动子,它们可在大肠杆菌中指导转录。当淋病奈瑟菌在正常生长条件下体外培养时,这些启动子似乎不会在可检测水平发挥作用。T7 RNA聚合酶表达系统已被用于鉴定该质粒编码的蛋白质,构建了一系列亚克隆并用于定位编码每种蛋白质的基因。在对序列数据库的搜索中,我们发现该质粒一个1.2 kb片段的推导氨基酸序列与大肠杆菌产大肠杆菌素质粒ColA、ColE1和ColK编码的Mob蛋白具有高度的序列相似性。1.2 kb片段包含Col转移区域的一部分,该区域随后因一个小的重复而失活。在形成隐匿质粒的重组事件中,其中一个转移基因与一个隐匿质粒开放阅读框融合,形成cppB基因。尽管cppB基因是通过这种方式形成的,但我们已表明,当淋病奈瑟菌在适当的环境条件下生长时,cppB基因可以表达。