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编码增强血清存活能力和表面排斥功能且与λDNA具有同源性的质粒决定因子iss的克隆及DNA序列分析

Cloning and DNA sequence of plasmid determinant iss, coding for increased serum survival and surface exclusion, which has homology with lambda DNA.

作者信息

Chuba P J, Leon M A, Banerjee A, Palchaudhuri S

机构信息

Department of Immunology and Microbiology, Wayne State University School of Medicine, Detroit, MI 48201.

出版信息

Mol Gen Genet. 1989 Apr;216(2-3):287-92. doi: 10.1007/BF00334367.

Abstract

Escherichia coli K12 cells carrying a cloned 1.4 kb HindIII fragment from plasmid ColV2-K94, showed increased survival in guinea pig serum. The recombinant plasmid also conferred group II surface exclusion, i.e. the cells were reduced in recipient ability towards the incoming plasmid R538drd in conjugation experiments. Southern blotting suggested homology with bacteriophage lambda DNA and to the insertion element IS2. Determination of the DNA sequence of the fragment demonstrated the presence of a truncated IS2 (165 bp), separated by 250 bp from a 900 bp stretch of homology with lambda DNA, beginning within the Rz gene and continuing in the rightward direction on the lambda map. A 97 amino acid open reading frame (ORF) adjacent to Rz and on the opposite strand, remained intact in iss, with several amino acid changes. The ORF in iss is preceded by sequences resembling prokaryotic ribosome binding sites and promoters.

摘要

携带从质粒ColV2-K94克隆的1.4 kb HindIII片段的大肠杆菌K12细胞,在豚鼠血清中的存活率有所提高。该重组质粒还赋予了II型表面排斥作用,即在接合实验中,细胞对导入的质粒R538drd的受体能力降低。Southern印迹分析表明,该片段与噬菌体λ DNA和插入元件IS2具有同源性。对该片段的DNA序列测定表明,存在一个截短的IS2(165 bp),与λ DNA的900 bp同源序列相隔250 bp,该同源序列从Rz基因内部开始,并在λ图谱上向右延伸。与Rz相邻且位于相反链上的一个97个氨基酸的开放阅读框(ORF)在iss中保持完整,但有几个氨基酸发生了变化。iss中的ORF之前有类似于原核生物核糖体结合位点和启动子的序列。

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