小鼠抗人小细胞肺癌单抗轻链可变区基因的体外扩增、克隆及序列分析

Cloning and sequencing of mouse immunoglobulin kappa variable domains against human small lung cancer by the polymerase chain reaction

本文通过一对分别位于小鼠免疫球蛋白轻链可变区基因 FR1骨架区和 FR4骨架区的通用引物,利用 PCR 技术首次从小鼠抗人小细胞肺癌2F7杂交瘤细胞染色体组总 DNA 中直接扩增出相应的免疫球蛋白轻链可变区基因,扩增出的基因片段经引物上引进的两个限制性内切酶位点直接克隆到装有人免疫球蛋白轻链恒区的克隆载体 pGEM-7Zf(+)-V_kPCR 中的 EcoR1和 BglⅡ内切酶位点之间.通过对其进行顺序分析,结果表明我们克隆得到了免疫球蛋白轻链可变区基因,其长度为318bp.

We have directly amplified immunoglobulin kappa variable domains from the genomic DNA of 2F7 ybridoma against human small cell lung cancer by the polymerase chain reaction(PCR)with a set of universal primers that were on the FR1 site and FR4 site of V_(?) genesrespectively.By EcoRI and BglⅡ,the DNA fragments amplified were force-cloned into pGEM-7Zf(+)-V_kPCR that carried human immunoglobulin kappa constant domain.By sequencing,It conformed that we had got the V_(?) gene that was 318 bp in length.