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一株尿路肺炎克雷伯菌1型菌毛重组质粒的构建与表达

Construction and expression of recombinant plasmids encoding type 1 fimbriae of a urinary Klebsiella pneumoniae isolate.

作者信息

Purcell B K, Clegg S

出版信息

Infect Immun. 1983 Mar;39(3):1122-7. doi: 10.1128/iai.39.3.1122-1127.1983.

Abstract

The type 1 fimbriae of Klebsiella pneumoniae have been implicated as important virulence factors in mediating Klebsiella urinary infections. The chromosomally encoded fimbrial genes were cloned by a cosmid cloning technique. Further subcloning was performed with the cloning vehicles pBR322 and pACYC184, and a recombinant plasmid containing the fimbrial genes was constructed. After transformation by this plasmid, both Escherichia coli and Salmonella typhimurium were shown to express fimbriae which reacted with Klebsiella fimbrial antiserum. The approximate location of the relevant genes on the chimeric plasmid was determined by insertion of the transposable element Tn5. Hemagglutination-negative phenotypes were used to estimate the minimum size of the DNA fragment necessary to encode fimbrial biosynthesis and expression. The size of the coding region of this fragment was found to be 5.5 kilobase pairs.

摘要

肺炎克雷伯菌的1型菌毛被认为是介导克雷伯菌泌尿系统感染的重要毒力因子。通过黏粒克隆技术克隆了染色体编码的菌毛基因。使用克隆载体pBR322和pACYC184进行了进一步的亚克隆,并构建了一个含有菌毛基因的重组质粒。用该质粒转化后,大肠杆菌和鼠伤寒沙门氏菌均显示表达能与克雷伯菌菌毛抗血清发生反应的菌毛。通过插入转座元件Tn5确定了嵌合质粒上相关基因的大致位置。利用血凝阴性表型来估计编码菌毛生物合成和表达所需的DNA片段的最小大小。发现该片段编码区的大小为5.5千碱基对。

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