肺炎支原体P1蛋白羧基端基因片段在大肠杆菌中克隆的研究

在大肠杆菌中克隆肺炎支原体P1蛋白羧基端基因片段,为P1蛋白基因片段的扩增、表达及探讨羧基端基因片段功能打基础.采用PCR扩增方法获取P1结构基因.扩增产物用SalI和EcoRI酶切消化,回收1kb大小的DNA片段并与pUC19DNA连接,转入大肠杆菌JM109菌株.用X-gal平板及质粒图谱分析方法筛选重组克隆株,再用限制性核酸内切酶酶切图谱分析鉴定.经PCR扩增MPDNA获得1条5.0kbDNA片段.重组质粒限制性内切酶指纹图谱显示出2条带,1条为pUC19载体DNA带,另1条是1kb的插入片段.实验获得肺炎支原体P1蛋白结构基因及含P1蛋白羧基端DNA片段的重组克隆株.

CLONING OF P1 PROTEIN C END GENE FRAGMENT OF Mycoplasma pneumonia INTO E.coli

Cloning of P1 protein C end gene fragment of M. pneumonia into E. coli laid a foundation of its amplification, expression, and probe into its function. The P1 structure gene was amplified by PCR method. The 1kb DNA fragments were recovered and linked with pUC19 vector DNA, then transformed into E. coli JM109. The recombinant clones was screened by X-gal plate and plasmid map analysis, and identified by restricted enzyme mapping analysis. A 5.0kb DNA fragment was obtained by MP DNA amplified with PCR. The fingerprint of endonuclease mapping of the recombinant plasmid showed two bands, one was pUC19 DNA band, and the other was 1kb insert fragment.