法氏囊B细胞λ轻链基因编码蛋白在大肠杆菌中的表达  

Expression of Protein Encoded by λ Light Chain Gene of Bursal B Lymphocytes in E.coli

作　　者：宋海涛[1] 张红[1] 王丽[2] 段艳华[1] 乔松林[1] 李学伍[2] 赵光辉[1] 张改平[2] 

机构地区：[1]河南农业大学,河南郑州450002 [2]河南省动物免疫学重点实验室,河南郑州450002

出　　处：《河南农业科学》2007年第3期109-111,共3页Journal of Henan Agricultural Sciences

基　　金：国家自然科学基金重点项目(30203270)

摘　　要：为获得鸡λ轻链基因的表达蛋白,采用聚合酶链反应从鸡法氏囊B细胞cDNA文库中扩增出分泌表达蛋白的编码基因;用限制性内切酶消化后,插入克隆载体pUC19中。经PCR鉴定与序列测定证实后,以亚克隆法构建于原核表达载体PET43a的相应酶切位点。经过基因序列测定、BamHⅠ和HindⅢ双酶切鉴定证实克隆载体正确插入载体PET43a。转化宿主菌BL21(DE3),经IPTG诱导、SDS-PAGE分析表明目的蛋白得到表达;且能与His-Taq单抗相结合。λ light chain gene encoding the expression protein was amplified by polymerase chain reaction（PCR） from infectious bursal B lymphocyte cDNA library, and inserted into the corresponding sites of vector pUC19 after digested by restriction endonuclease to construct protocaryon expression vector PET43a. Both nucleotide sequencing and BarnH Ⅰ and Hind Ⅲ digestion confirmed that λ light chain gene fragment encoding the target protein was correctly inserted into the vector. The recombinant plasmid was transformed into E. coli BL21 （DE3） and induction by IPTG and SDS-PAGE indicated that the target protein was expressed.

关 键 词：传染性法氏囊 基因 蛋白 表达 

分 类 号：S858.31[农业科学—临床兽医学]