触减PCR在扩增抗体重链和轻链可变区中的应用  

Application of touch-down PCR for amplifying heavy-chain and light-chain variable region genes of immunoglobulin

作　　者：秦琴[1] 刘建荣[1] 王毅民[1] 董丽娜[1] 高嵩丹[1] 

机构地区：[1]山西省人民医院中心实验室,太原030012

出　　处：《山西医科大学学报》2010年第1期91-93,共3页Journal of Shanxi Medical University

基　　金：山西省人民医院青年科研基金资助项目(200803)

摘　　要：目的探讨扩增人免疫球蛋白重链和轻链可变区基因的方法。方法提取经免疫的具有乙肝表面抗体的人外周血淋巴细胞总RNA,反转录合成cDNA,采用普通PCR和触减PCR两种方法扩增抗体重链和轻链可变区基因。结果利用触减PCR方法,扩增出抗体重链和轻链的可变区基因,基因片段分别约为340bp和325bp,而普通PCR没有得到预期的结果。结论触减PCR比普通PCR更容易扩增出抗体可变区基因片段。Objective To explore a better method to amplify heavy-chain and light-chain variable region genes（ VH and VL）of immu- noglobulin. Methods Total RNA was extracted from human peripheral blood lyrnpbocytes（PBL） from healthy volunteer with antibody against HBsAg,and converted into eDNA by reverse transeriptase. The variable region genes（VH and VL）of immunoglobulin were am- plified by ordinary PCR and touch-down PCR（TD PCR）. Results VH and VL genes were successfully amplified by TD PCR,and the lengths were 340 bp and 325 bp respectively. However, VH and VL genes were not amplified by ordinary PCR. Conclusion TD PCR is a better method to amplify the VH and VL gene than ordinary PCR.

关 键 词：触减PCR 噬菌体展示技术 抗体库 

分 类 号：R-33[医药卫生]