关键词: 胶质瘤;噬菌体抗体库;单链抗体;基因

    中图号:R392.11

    摘 要:目的 构建人源性胶质瘤噬菌体抗体库. 方法 从胶质瘤患者外周血淋巴细胞中提取细胞总RNA，经逆转录后用套式PCR分别扩增抗体轻链Vκ 和重链VH 基因.分别构建Vκ 5.5×106 ，VH 3.5×106 基因库，连接Vκ 和VH 构建单链抗体(ScFv)基因库，并克隆入表达载体pDNA5内进行重组，得到ScFv噬菌体抗体库. 结果 表明经Cre-Loxp511系统重组后，噬菌体抗体库库容量达到6.0×108 ，测序证实抗体基因与人源性抗体基因数据库同源. 结论 利用Cre-Loxp511胞内重组系统成功地构建了较大容量的人源性脑胶质瘤噬菌体抗体库，为进一步筛选特异性人源性胶质瘤ScFv奠定了基础.

    Construction of phage library of human glioma ScFv antibody by Cre┐Loxp recombinaton system

    WANG Yan-Gang，ZHANG Xiang，FEI Zhou，LI Bing，GUO Qing-Dong，LIU Xian-Zhen，ZHANG Yong-Qin，ZHAO AI-Zhi

    Institute of Neurosurgery of Chinese PLA

    Department of General Surgery，Xijiing Hospital，Fourth Military Medical University，Xi'an710033，China

    Keywords:glioma;phage antibody library;single chain Fv;gene

    Abstract:AIM To construct recombination library of hu-man glioma ScFv antibody by phage display technology.METHODS Total RNA was extracted from peripheral blood lymphocytes of patients with glioma and reversely transcript-ed into cDNA.Variable region genes(Vκ and VH gene)were amplified from the cDNA using nested polymerase chain reac-tion.Vκ (3.5×106 )and VH (1.5×106 )gene library was first-ly constructed and Vκ gene with VH gene were connected ......