特异性抗IBDV单链抗体基因的构建及序列分析  

Construction and sequencing of variable region genes of the single strand antibody recognizing IBDV

作　　者：胡昕[1] 陆苹[1] 李晖[1] 孙建和[1] 王国丰[1] 

机构地区：[1]上海交通大学农学院,上海201101

出　　处：《上海交通大学学报（农业科学版）》2002年第1期58-62,共5页Journal of Shanghai Jiaotong University(Agricultural Science)

摘　　要：从抗 IBDV杂交瘤细胞系 SJS中分离总 RNA,经 RT-PCR体外扩增重、轻链可变区基因 ,通过一连接肽 (Giy4 Ser) 3基因拚接形成 Sc Fv基因 ,经 IBDV抗原亲和筛选出特异性阳性克隆进行核苷酸序列测定及氨基酸序列推导。所获得的特异性抗 IBDV单链抗体基因为 VH-Linker-VL 结构 ,其含有编码 (Giy4 Ser) 3的连接肽基因及维持抗体结构所必须的半胱氨酸残基 ,编码产物具有与亲本抗体相同的抗原结合活性和特异性 ,从而为研制具有应用潜力的高表达工程化抗体奠定了基础。To obtain the genes of a single chain antibody from mAb SJS against IBDV,The gene of variable region of heavy and light chain were amplified separately by RT PCR from the hybridoma cell line secreting mAb SJS and assembledinto ScFv gene with a (Gly 4Ser) 3 linker by the splicing overlap extension(SOE).The IBDV specific ScFv gene was sequenced after affinitive selection.The results showed that the fusion gene which was capable of encoding affinitive and specific product against IBDV consist of the variable regions of light and heavy chain in mouse anti IBDV antibody and a 45bp Linker .The constructed ScFv gene is a functional ScFv of the specific mAb SJS against IBDV.

关 键 词：传染性法氏囊病病毒 可变区 单链抗体 噬菌体展示 基因克隆 序列分析 RT-PCR 鸡 

分 类 号：S858.315.3[农业科学—临床兽医学] S858.312.5[农业科学—兽医学]