抗鹅细小病毒噬菌体轻链抗体文库的构建  

Construction of specific light chain of phage antibody library of anti-goose parvovirus

作　　者：吕雪峰[1] 李志慧[2] 刘钧松[2] 呼延含蓉 杨金生[1] 王茹丽 

机构地区：[1]吉林省畜牧兽医科学研究院,吉林长春130062 [2]吉林大学超硬材料国家重点实验室,吉林长春130012

出　　处：《中国兽医学报》2010年第11期1432-1434,共3页Chinese Journal of Veterinary Science

基　　金：家畜疫病病原生物学国家重点实验室开放式基金资助项目(08002);吉林省科技发展计划资助项目(20090162)

摘　　要：取鹅细小病毒(Goose parvolirus,GPV)免疫过的抗体阳性的试验鹅的脾脏,分离淋巴细胞,提取细胞总RNA后反转录,利用鹅IgG基因特异性引物,分别扩增轻链基因。将鹅轻链基因克隆入噬菌体载体pComb3,经电穿孔法转化大肠杆菌XL1-Blue,再以辅助噬菌体VCSM13超感染,构建抗GPV噬菌体轻链抗体库,筛选阳性克隆,测序分析。结果显示,构建了容量为1×106、重组率为66.6%的轻链抗体库。The total RNA was extracted from spleen lymphocyte of the goose which immuned by goose parvovirus.Light-chain segment of goose IgG gene were amplified respectively by RT-PCR using a set of specific primers.The amplified gene were inserted successively into vector pComb3 and electrotransformed E.coli XL1-Blue furthermore,the recombinant phage was rescued by being concultured with helper phage VCSM13 to construct specific phage antibody library,sequence and analyze.The protein was displayed and constucted 1×106 clone library was established.The recombination fraction is 66.6%.

关 键 词：鹅 细小病毒 噬菌体展示 轻链 

分 类 号：S852.65[农业科学—基础兽医学]