系列鼠抗CD分子单克隆抗体轻、重链可变区基因的克隆和序列分析

目的 获取系列鼠抗人CD分子单抗的轻链及重链可变区基因并进行克隆和测序。方法 采用RT－PCR技术，从WuT4、WUT3、WuTac、WuLCA、WuT9杂交瘤细胞总RNA中分别扩增出轻链和重链可变区基因，进行克隆并作核苷酸序列分析。结果 以上几种抗体的轻重链可变区的 PCR产物大小均为 400bp左右，成功构建了pMD18－T－VL和 pMD18－T－VH克隆载体，经限制性酶切分析，其大小与预期结果相符，并按ABI PRLSM BigDye系统测序。结论 经与Kabat、Blast抗体基因同源数据库比较，所获基因片段均为抗体基因。

Cloning and Sequencing of VH and VL Variable Region Genes of Series Murine McAbs to Human CD Molecule

[Abstract] Objective To obtain the VH and VL variable region genes of murine McAbs to human CD molecule. Methods The VH and VL variable region genes were amplified from the total RNA of WuT3, WuT4, WuTac, WuLCA and WuT9 hyridoma cells by RT - PCR separately, then cloned and sequenced. Results All the length of the PCR products were about 400bp. Restriction enzyme analysis proved that cloning vectors pMDIS - T - VL and pMDIS - T - VH were successfully constructed, and the sequencing results of them were in accord with expected. Conclusion All the obtained fragments were proved to be ahtibody genes in comparison with Kabat and Blast antibody gene homology database.