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

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.