牛泡沫病毒BFV3026 gag基因的克隆及其在大肠杆菌中的表达(英文)  

CLONING OF BFV3026 GAG GENE AND EXPRESSION IN E.coli

作　　者：余荭[1] 孔晓红[1] 马永刚[1] 李汀[1] 王金忠[1] 陈启民[1] 耿运琪[1] 

机构地区：[1]南开大学生命科学学院,天津300071

出　　处：《南开大学学报（自然科学版）》2003年第1期84-90,共7页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：Supported by National Science Foundation of China(C3 9970 0 3 3 )

摘　　要：从牛泡沫病毒 BFV3 0 2 6感染的胎牛肺细胞中提取 Hirt DNA,PCR扩增 BFV3 0 2 6gag基因 .序列分析确证后 ,将其克隆于原核表达载体 p ET3 2 A,重组质粒转化大肠杆菌 BL2 1 (DE3 ) ,经 IPTG诱导 ,使 Gag蛋白得以高效表达 .SDS-PAGE及 Western blot证实 :表达蛋白占菌体总蛋白的 40 % .本工作的完成为 Gag蛋白功能的研究奠定基础 .With polymerase chain reaction (PCR), a fragment encoding the Gag protein was amplified from the Hirt DNA of bovine foamy virus3026-infected cells, which was isolated previously from the peripheral blood mononuclear cells of an infected cattle in Tianjin.. Sequence analysis revealed that it had high identity with that of the reported BFV in GenBank. Then this gag gene was ligated into plasmid of pET 32A for prokaryotic expression. BL21 (DE3) of E.coli transformed with the recombinant plasmid of pET gag was induced to specifically express the Gag protein in high level. The expressed product was characterized by SDS-PAGE and Western blot analysis, which occupies 40% of total bacterial protein. The report has laid a basis for searching the possible function of Gag in virus assembly.

关 键 词：牛泡沫病毒 BFV3026 GAG基因 基因克隆 基因表达 序列分析 大肠杆菌 

分 类 号：S852.653[农业科学—基础兽医学] Q78[农业科学—兽医学]