抗膀胱癌单克隆抗体重链可变区基因扩增克隆及序列测定

从分泌人膀胱癌特异性单克隆抗体的杂交癌细胞ＢＤＩ－１中提取总ＤＮＡ，应用人工合成的引物经聚合酶链反应（ＰＣＲ）扩增出重链可变区基因片段，将扩增产物插入ＰＵＣ１９质粒，筛选出阳性克隆，用链终止法进行ＤＮＡ序列测定分析，结果证实抗体重链可变区基因片段全长由３６６个碱基组成。

Amplification cloning and sequencing of immunoglobulin heavy variable-region gene of the monoclonal antibody anti-bladder carcinoma

Total DNA was extracted from a cultured hybridoma cell line BDI-l,which secretes human bladder carcinoma specific monoclonal antibody.The variable region gene of immunoglobulin heavy chain was amplified from the DNA with pairs of synthetic oligomers as primers by polymerase chain reaction (PCR).The PCR product was ligated to PUC19.By screening and identification,several recombinants that had been inserted with the target fragments were obtained.The recombinants were sequenced by Sanger's method.Our result shows that the length of heavy variable-region gene of this antibody is 366 base pairs.