含有弓形虫表面抗原P22、P30复合基因的载体构建  

Construction of P30 and P22 compound gene of the major surface antigen of Toxoplasma gondii

作　　者：王伟[1] 古钦民[2] 刘师莲[1] 何深一[2] 胡海燕[1] 李瑛[2] 卢翌[1] 耿昭[1] 

机构地区：[1]山东大学医学院医学分子生物学实验中心,济南250012 [2]山东大学医学院寄生虫学教研室

出　　处：《中国人兽共患病杂志》2002年第6期43-45,共3页Chinese Journal of Zoonoses

基　　金：山东省卫生厅资助项目 (99CA1CAA10 )

摘　　要：目的　选择弓形虫表面抗原P2 2、P30的有效基因片段 ,共同构建在同一克隆载体pUC19和表达载体 pGE MEXTM- 1上 ,并保证其连接方向及开放读码框的正确。方法　根据已发表P30基因序列 ,用PCR技术调取所需基因片段 ,P2 2基因片段来自重组质粒pGEX - 2T -v2 2 ,将两片段定向克隆于克隆载体 pUC19及表达质粒pGEMEXTM- 1上 ,用酶切及测序的方法对重组子进行鉴定。结果　酶切产物经电泳显示条带清晰 ,P2 2、P30基因片段的泳动位置分别在 4 4 6bp和 86 0bp的位置 ,与预计结果一致 ;测序结果表明插入的复合基因片段方向及序列均正确。结论　成功构建的含有P2 2、P30基因片段的质粒重组体 ,保证了两基因连接方向、序列及开放读码框的正确 ,为今后两基因的进一步复合表达研究奠定了基础。Aim To select the major surface antigen P22,P30 of Toxoplasma gondii,and insert them into the pUC19 and pGEMEX TM 1 correctly Methods A pair of primer were designed and synthesized according to the published gene sequence of the p30 gene,by enzyme cutting and measuring sequence to ensure the correct of the recomposed Result The enzyme cutting of the recomposed plasmid showed that the fragments of P22 gene and P30 gene were 446bp and 860bp,the measuring showed the sequence was correct too Conclusion The clone plasmid (pUC19 p22 p30)and expression plasmid (pGEMEX TM p22 p30) have been established successfully,reassuring the precision of open reading framework

关 键 词：弓形虫 P30基因 P22基因 克隆 疫苗 

分 类 号：R531.8[医药卫生—内科学]