摘要
目的克隆ＨＡｂ２５肝癌单克隆抗体（ＭＣＡｂ）的可变区基因。方法ＲＴ－ＰＣＲ＄从分泌ＨＡｂ２５单克隆抗体的杂交瘤细胞中，扩增出ＶＨ和ＶＬ基因，双脱氧链终止法测定其核苷酸序列。结果ＶＨ和ＶＬ基因的两端均含有完整的引物序列，ＶＨ基因全长３６０ｈＰ，编码１２０个氨基酸，ＶＬ基因全长３３０ｈＰ，编码１１０个氨基酸，重、轻链可变区内均仅含单一开放读框，具有明显的抗体可变区特征，与基因数据库（ＧｅｎＢａｎｋ冲的序列进行比较分析，基因库中无相同的基因，ＶＨ基因与小鼠重链ＶＨ１８６．２家族同源性最高（８４．００％），属小鼠屹重链可变区９个家族中的第三族，ＶＬ基因与小鼠ｋａｐｐａ轻链ＭＭＩｇＧＫＡＶＡＧ基因同源性最高（８２．００％），属小鼠Ｉｇｋａｐｐｐａ轻链第ＩＶ组。结论克隆的ＨＡｂ２５ＭｃＡｂ的可变区基因为功能性重排的小鼠可变区基因。
Objective To clone the variable gmes of HAb25 McAb against hepatocellular carcinoma.Methods The HAb25 variable region genes were isolated by RT-PCR technique from the HAb25hybridoma, and its nucleic acid opuences were analyzed by the sandals dideoxy-mediated chain-termi-nation method. Besults The gene of VH was 360bp long encoding 120 amino acids and the gene of VLwas 330bp long encoding 110 amino acids. Only one open reading frame could be found in each of thevariable genes. A comparison of the opuences of VH and VL domatins derived from the HAb25 antibodywith those of the published mouse ig genes in GenBank by computer revealed that the VH bine washomologous to the VH186.2 g6rmline g6ne family (84.00%) and VL gene was homologous to theMMIgGKAVAG(82.00%). Conclusion Both of the two genes are rearranged varisble region genes, whichare VHDJH3 for the VH and VK J K4 for the VL.
出处
《中华肝脏病杂志》
CAS
CSCD
1999年第2期101-103,共3页
Chinese Journal of Hepatology
