SOE PCR重构SLA-Ⅰ复合体研究  

Study on Reconstruction of SLA-Ⅰ Complex by SOE PCR

作　　者：陈燕宇[1] 何丹[1] 陈兆华[1] 周巧妮[1] 马文军[2] 高凤山[1] 

机构地区：[1]大连大学生物工程学院,辽宁大连116622 [2]河北省秦皇岛市海港区畜牧局,河北秦皇岛102200

出　　处：《动物医学进展》2010年第9期58-62,共5页Progress In Veterinary Medicine

基　　金：大连大学本科生创新教育基金项目(2009244)

摘　　要：利用剪接重叠延伸PCR(SOE PCR)技术,中间加一富含甘氨酸/丝氨酸的linker(G4S)3,将SLA-Ⅰ重链基因SLA-2和轻链基因β2m连接,形成复合体基因链SLA-2-(G4S)3-β2m,然后将复合体基因链克隆入p2X表达质粒,导入受体菌TB1并进行表达。结果显示,利用SOE PCR技术成功得到了复合体基因链SLA-2-(G4S)3-β2m,并且克隆入p2X载体。SDS-PAGE检测表明,复合体基因导入宿主菌后获得了融合蛋白MBP-SLA-2-(G4S)3-β2m,大小为84.1 ku。试验结果为今后在体外进行相关基因重组表达提供参考。By SOE PCR,the SLA-2 gene derived from SLA-Ⅰ heavy-chain molecule along with the light-chain gene β2m were linked to be a combined gene that named as SLA-2-(G4S)3-β2m,via a linker(G4S)3 encoding glycine/serine rich sequence.Then the combined gene SLA-2-(G4S)3-β2m was cloned into the expressing vector p2X followed by transforming the recombinant vector into E.coli TB1 and inducing them to be expressed.The result indicated that the combined gene SLA-2-(G4S)3-β2m was obtained by SOE PCR and cloned into p2X successfully.After being transformed into TB1 and induction,the combined gene was expressed to be a fusion protein MBP-SLA-2-(G4S)3-β2m with 84.1 ku.This research will supply a reference to recombine and express some relative genes in vitro.

关 键 词：剪接重叠延伸 PCR SLA-Ⅰ 复合体 

分 类 号：Q78[生物学—分子生物学]