日本血吸虫(中国大陆株)22.6 ku抗原编码基因的核酸疫苗pCMV/Sj22.6的构建

Constraction of pCMV / Sj22.6 Naked DNA Vaccine of Schistosoma japonicum (Chinese Mainland Strain)

目的 探索日本血吸虫 (中国大陆株 ) 2 2 .6ku抗原 ( Sj2 2 .6)编码基因用作核酸疫苗的可行性。方法 用新设计的引物以 PCR法对 Sj2 2 .6编码基因进行改造 ,然后亚克隆入真核表达载体 p CMV -β并转化入大肠杆菌 JM 10 9。结果 提取重组质粒经酶切分析 ,筛选出了含有正向插入片段的阳性重组质粒。

Objective To investigate if the gene fragment of 22.6 ku antigen of Schistosoma japonicum (Sj22.6) could be expressed in eukaryotic cells. Methods It was reformed by PCR to introduce Not Ⅰ sites at both ends, a Kozak sequence at the 5′ end and remove stop coden TAA ahead of the first start coden ATG. Then it was subcloned into eukaryotic plasmad pCMV β and transformed into E.coli JM109. Results The Positive clone containing the correct orientation recombinant pCMV / Sj22.6 was picked out. Conclusion It could be used in further studies.