免疫佐剂分子CTB基因的克隆与表达

Cloning and Expression of Immunoadjuvant Molecule-CTB Gene

我们采用聚合酶链式反应从霍乱弧菌 5 6 9B和 M0 4 5中扩增得到霍乱弧菌 B亚基基因 ,导入载体p UC18,构建重组质粒 p CTB并转化大肠杆菌 JM10 9,并用限制性酶切分析、聚合酶链式反应、序列分析、硫酸十二烷酸钠 -聚丙烯酰胺凝胶电泳、Western印迹进行鉴定。实验结果表明我们成功地扩增出 376 bp的霍乱弧菌 B亚基基因 ;构建免疫佐剂分子的重组质粒 p CTB;并在原核系统中表达出 12 KD的霍乱弧菌 B亚基抗原区带。

We cloned cholera toxin subunit B gene from 569B and M045 strain of Vibrio cholerae with polymerase chain reaction, constructed recombinant plasmid pCTB, and transformed pCTB into the prokaryotic cell strain JM109. The indentification was made by means of restriction enzyme analysis, polymerase chain reaction, DNA sequencing, SDS polyacrylamine gel electrophoresis analysis and Western blot. The results indicate that we have amplified cholera toxin subunit B gene of 376bp from Vibrio cholerae and hve constructed the recombinant plasmid pCTB, and we have affained the object amied at successful expression of 12KD in the prokaryotic cell strain.