5’-RACE法扩增抗人乳头瘤病毒16型L1蛋白单克隆抗体轻链可变区基因及序列分析  

Amplifying variable region gene of light chain （VL） of monoclonal antibody against human papillo- mavirus 16 L1 （ HPV16 L1 ） protein by 5＇ -RACE and sequence analysis

作　　者：王燕[1] 商庆龙[1] 陈思佳[1] 邵迪[1] 韩聪[1] 李茉[1] 谷鸿喜[1] 

机构地区：[1]哈尔滨医科大学微生物学教研室,黑龙江省普通高校病原生物学重点实验室,150081

出　　处：《国际免疫学杂志》2008年第4期247-250,共4页International Journal of Immunology

基　　金：黑龙江省科技攻关项目（GB02C111）;哈尔滨市科委重大攻关项目（2004AA3CS176-1）;黑龙江省青年科学技术专项资金项目（QC06C062）;黑龙江省卫生厅重点项目（2006-253）

摘　　要：目的获得抗人乳头瘤病毒16型（HPV16）L1蛋白单克隆抗体的轻链可变区（VL）基因并分析序列。方法从分泌抗HPVl6L1蛋白单克隆抗体的杂交瘤细胞中提取总RNA，逆转录形成CD-NA，用5’-RACE策略扩增抗体轻链可变区基因，经琼脂糖凝胶电泳鉴定，并测序及进行序列分析。结果VL基因全长336bp，编码112个氨基酸，基因测序结果符合小鼠抗体轻链可变区特征。结论5’-RACE法成功获得了抗HPV16L1蛋白的单克隆抗体轻链可变区基因的真实序列，为基因工程抗体研究奠定了良好基础。Objective To acquire the variable region gene of light chain （VL） of monoclonal antibody against human papillomavirus 16 L1 （ HPV16 L1 ） protein. Methods Total RNA was extract from hybridoma cells secreting specific monoclonal antibody against HPV16L1, transcripted reversely into cDNA with random primers. The variable region of the light chain gene fragments was ampliflied using 5＇ -RACE. Sequencing was confirmed by agarose gel electrophoresis and sequencing analysis. Results Full length of VL gene was 336bp that encoding 112 amino acids. The VL gene was homologous with the published gene sequences of mouse antibody variable region. Conclusion The light chain sequence of monoclonal antibody against HPV16L1 protein was obtained by 5＇ -RACE, which provides a good basis for construction of a recombinant antibody against HPV16 L1.

关 键 词：人乳头瘤病毒16型 轻链 可变区 单克隆抗体 

分 类 号：R73[医药卫生—肿瘤] R3[医药卫生—临床医学]