抗人CD3单抗轻、重链可变区基因的克隆及序列分析  

Cloning and Sequence Analyses of the V H and V K Genes of an Anti human CD3 Monoclonal Antibody

作　　者：萧飒[1] 顾征[1] 王勇[1] 陈艾[1] 林晴[1] 张卫国[1] 蒋欣[1] 黄华梁[1] 沈倍奋 

机构地区：[1]中国科学院遗传研究所

出　　处：《生物工程学报》1996年第S1期121-125,共5页Chinese Journal of Biotechnology

摘　　要：根据抗体基因可变区两端保守序列ＦＲ１和ＦＲ４，设计并化学合成轻重链ＰＣＲ扩增引物。从体外培养细胞ＵＣＨＴ１中提取总ＲＮＡ，反转录成ｃＤＮＡ，以ｃＤＮＡ为模板经ＰＣＲ扩增轻，重链可变区基因片段。将扩增片段克隆至ｐＵＣ１９质粒，筛选阳性克隆并测序，序列分析结果为重链３６６ｐｂ，编码１２２个氨基酸，轻链３２７ｂｐ。The two sets of PCR primers were designed based on the conserved sequences FR1 and FR4 of antibody variable genes.Hybridoma cell line UCHT1 were cultured,and the total RNA were extracted.By reverse transcriptation,the cDNA were synthesized and used as templates for PCR,and the desired V H and V K gene fragments were ampified.The PCR products were cloned into pUC19,and the recombinants were screened and sequenced.The sequence analyses results showed that V H is in length of 366 bases for 122 amino acids and V K is 327 bases for 109 amino acids.

关 键 词：UCHT1 抗体可变区基因 PCR 序列分析 

分 类 号：Q344.1[生物学—遗传学]