抗日本血吸虫卵单抗33F5可变区轻、重链基因的串联及克隆

LIGATION AND CLONING OF THE VARIABLE REGION GENES OF ANTI-SCHISTOSOMA EGG MONOCLONAL ANTIBODY 33F5

目的 制备抗日本血吸虫虫卵尿素溶性抗原单克隆抗体 3 3 F5的基因工程单链抗体。方法 利用杂交瘤细胞总 m RNA抽提、逆转录技术及 PCR方法、获得单抗重链和轻链可变区基因 ,通过linker将其连接 ,并克隆到表达载体 PCANTAB5 E中。结果 酶切显示 ,重链和轻链可变区基因及Sc Fv片段与预设计的大小一致。结论 获得抗日本血吸虫卵单抗 3 3 F5可变区轻。

Objective To prepare the genetic engineering antibody of the variable region gengs of anti Schistosoma egg monoclonal antibody 33F5. Methods mRNA was extracted from 33F5 hybridoma cells. cDNA fragments for heavy and light variable regions were amplified by reverse transcription polymerase chain reaction. The amplified VH and VL were ligated into single fragment(ScFv) by linker, and then ligated fragment was cloned into expression plasmid PCANTAB5E. Results The recombinant plasmid was digested,the size of VH?VL and ScFv fragments were 340 bp?320 bp and 750 bp respectively . Conclusion The recombinant plasmid of anti Schistosoma egg monoclonal antibody 33F5 was constructed.