摘要
应用RT_PCR技术,从分泌具有中和活性的抗A型产气荚膜梭菌α毒素单克隆抗体(McAb) 的杂交瘤细胞1A8 中,扩增出抗体VH 和VL基因,用linker(Gly4Ser3)3 基因,将VH 和VL 基因连接成ScFv 基因,并将其克隆至pGEM_T载体中。经核甘酸序列分析证实,VH 和VL 基因及linker 基因拼接正确,基因全长729bp,经计算机分析,VH 和VL基因均为新发现的基因序列,符合功能性重排的鼠抗体可变区基因特征。VH 和VL 基因分别属于鼠免疫球蛋白重链Ⅱ(A)
The V H and V L genes were amplified from a hybridoma cell line 1A8 producing mouse McAb against alpha_toxin of Clostridium perfringens type A by RT_PCR.The V H and V L genes were connected through a flexible linker(Gly 4Ser) 3 and the V H_linker_V L(ScFv)fusion gene was cloned into a clone vector pGEM_T.The ScFv gene was sequenced, analyzed by computer. The ScFv gene consists of 729bp encoding 243 amino acid residues. Both V H and V L genes were confirmed as functionally rearranged mouse immunoglobulin variable region genes and appeared to be new genes. According Kabat classed method, McAb 1A8 V H gene segment and V L gene segment belong to the mouse lg heavy chain subgroup Ⅱ(A) and κ chain subgroup Ⅵ respectively.
出处
《中国预防兽医学报》
CAS
CSCD
2000年第1期30-33,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
全军医药卫生科研基金!课题
98Q076
关键词
A型
产气荚膜梭菌
Α毒素
单链抗体
基因克隆
Clostridium perfringens type A
alpha_toxin
ScFv
gene cloning
nucleotide sequencing
