人源性丙肝病毒抗体轻链基因的克隆及重组表达载体的构建

Cloning of Gene Encoding Light Chain of Humanize Antibody Against Hepatitis C Virus and Construction of the Recombinant Expression Vector

目的构建人源性丙肝病毒抗体轻链基因表达载体。方法通过RT-PCR从丙肝病毒抗体强阳性的志愿者的外周血淋巴细胞中提取RNA,PCR扩增出人IgG轻链基因,将其克隆入噬菌粒pCom b3中,构建人源性丙肝病毒抗体轻链表达载体,并利用相应的方法对噬菌体抗体库进行初步鉴定。结果成功地构建了人源性丙肝病毒抗体轻链表达载体,酶切鉴定结果重组率达到88.8%。结论人源性丙肝病毒抗体轻链基因表达载体是高效的,为下一步构建丙肝病毒抗体F ab段基因重组载体奠定了基础。

Objective To construction the recombinant expression vector of gene encoding light chain of humanized antibody against hepatitis C virus （HCV）. Methods The total RNA extracted from PBLs of volunteers whose antibody in their blood is positive ,and light chain genes of the immunoglobulin were amplified by PCR. Then clone them into pComb3 vector and construct the recombinant expression vector. Results Constructed the recombinant expression vector of gene encoding light chain of humanized antibody against HCV successfully. The recombinant ratio of light chain was 88. 8% knowing from the result of restriction enzyme digestion and the result of PCR. Conclusion The expression vector of humanized antibody light chain gene of against HCV is constructed effectively which laya solid foundation for further constructing the recombinant expression vector of humanized antibody Fab fragment against HCV.