重症肌无力抗乙酰胆碱受体单链抗体基因的构建.PDFVIP

Journal of Medical Science Yanbian University 　Sep. 2004 Vol. 27 No. 3 1·62 · 重症肌无力抗乙酰胆碱受体单链抗体基因的构建 金桂花 ,王玉祥 ,范华英 ,李红花 ,魏成淑 ,李英信 ,孟繁平 (延边大学医学院 微生物学与免疫学教研室 ,吉林 延吉 133000) [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 369 bp ,轻链可 变区基因长度为 321 bp ,单链抗体基因长度为 735 bp ;单链抗体 A 7 基因正确地插入在载体质粒p HEN 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. [ 关键词] 　重症肌无力 ;受体 ,胆碱能 ;基因重组 ( ) [ 中图分类号] 　R 392 1 　　　[文献标识码] 　A 　　　[文章编号] 2004 Construction of gene of single chain variable fragment against acetylcholine receptor in myasthenia gravis JIN Guihua , WANG Yuxiang , FAN Huaying , L I Honghua , WEI Chengshu , L I Yingxin , MENG Fanping ( ) Depart ment of Microbiology and Im m unology , Yanbian U niversity College of Medicine , Yanji 133000, Jilin , China ABSTRACT : OBJ ECTIV E To construct the gene of single chain variable fragment ( ScFv) A 7 against the main immunogenic region ( M IR ) in acetylcholine receptor ( AChR ) . M ETHODS Using PCR and gene cloning technology , it was cloned that the heavy chain variable region (V H) and light chain variable region (VL ) genes. The constructed ScFv A 7 was checked in sequencing. R ESUL TS The sequencing showed that the size of the V H 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 p HEN 2. CONCL USION A ScFv A 7 gene against the M IR of