日本血吸虫(中国大陆株)Sj16基因的体外扩增及克隆

Amplification and Cloning of Sj16 gene of Schistosoma japonicum Chinese strain

为了构建日本血吸虫（中国大陆株）Ｓｊｌ６基因原核表达重组质粒ｐＧＥＸ－４Ｔ－Ｓｊ１６。根据曼氏血吸虫Ｓｍ１６基因已知序列设计合成一对引物．用 ＰＣＲ技术从日本血吸虫（中国大陆株）成虫ｃＤＮＡ文库中扩增 Ｓｊ１６基因；将Ｓｊ１６基因定向克隆人 ＰＧＥＸ－４Ｔ－１，转化感受态 ＢＬ２１／ＤＥ３菌；用酶切、ＰＣＲ扩增鉴定筛选得到的重组阳性克隆。结果表明，从日本血吸虫（中国大陆株）成虫ＣＤＮＡ文库中获取Ｓｊ１６基因，重组质粒中含有Ｓｊ１６基因。结果提示，成功构建日本血吸虫（中国大陆株）Ｓｊ１６基因原核表达重组质粒ｐＧＥＸ－４Ｔ－１－Ｓｊ１６，为进一步研究Ｓｊ１６基因功能打下基础。

To construst a prokaryotic expression plasmid contain a Sj 16 gene of Schistosoma japonicum Chinese strain,a couple of primers were designed according to the known sequence of Sm16 and the Sj16 gene obtained by amplification from cDNA library of Schistosoma japonicum Chinese strain by using PCR technique. By cloning Sj16 gene into a prokaryotic expression vector, pGEX-4T- 1, a recombinant plasmid pGEX-4T-1- Sj 16 was constructed and transferred into HL21 /DE3. The positive recombinant pGEX- 4T-1-Si16 was screened and identified by agarose gel electrophoresis endonuclease digestion and PCR technique. The results showed that the specific fragment of Sj16 was amplified and then the recombinant prokaryotic expression plasmid pGEX- 4T-1-Sj 16 was construsted by directional cloning. The results suggested that the recombinant prokaryotic expression plasmid containing Si 16 was successfully constructed, and it has given the basic material for studying the function of Sj 16 gene.