【摘要】 [目的 ]构建重症肌无力抗乙酰胆碱受体主要免疫原区单链抗体A 7基因 .[方法 ]应用PCR技术及基因克隆技术分两步完成重链和轻链可变区基因的克隆 ,再对构建的单链抗体A 7基因进行序列测定检测其核苷酸序列 .[结果 ]构建的重组质粒经序列分析 ,重链可变区基因长度为 36 9bp ,轻链可变区基因长度为 32 1bp ,单链抗体基因长度为 735bp ;单链抗体A 7基因正确地插入在载体质粒pHEN 2开放读码框架内 .[结论 ]成功地构建重症肌无力抗乙酰胆碱受体主要免疫原区单链抗体A 7基因 ,为进一步制备基因工程抗体奠定了基础 .更多还原

【Abstract】 OBJECTIVE?To construct the gene of single chain variable fragment (ScFv) A?7 against the main immunogenic region (MIR) in acetylcholine receptor (AChR).?METHODS?Using PCR and gene cloning technology, it was cloned that the heavy chain variable region (VH) and light chain variable region (VL) genes. The constructed ScFv A?7 was checked in sequencing.?RESULTS?The sequencing showed that the size of the VH gene, VL gene and the ScFv gene were 369?bp, 321?bp and 735?bp respectively; ScFv A?7 gene was correct and cloned into the open reading frame in pHEN?2.?CONCLUSION?A ScFv A?7 gene against the MIR of AChR has been successfully constructed.更多还原