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

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