摘要
将通过PCR技术改建并去除了信号肽的猪生长激素cDNA基因克隆入转移载体pBacPAK His1,构建了重组转移载体pPGH0 30 ,进而将pPGH0 30与线性化病毒Bm BacPAK6共转染BmN细胞 ,成功获得了猪生长激素重组杆状病毒Bm BacPAK6 pgh。感染重组病毒的细胞可溶性蛋白SDS PAGE和Western blot分析结果显示 ,感染细胞蛋白电泳带的 2 5kD处有 1条猪生长激素特异带。Bm BacPAK6 pgh的BmN细胞培养液接种家蚕 5龄幼虫和蛹 ,感病幼虫与蛹血淋巴的SDS PAGE和Western blot分析结果表明 ,6×His 猪生长激素融合基因在家蚕体内得到了表达 ,特异性条带大小约为 2 5kD。
The cDNA of pGH without signal peptides was reconstructed by PCR, then recombined with the vector pBacPAK-His1.The recombined plasmid,pPGH030,and linear Bm-BacPAK6 co-transfacted in the cell line of Bombyx mori (BmN),then the recombinant virus, Bm-BacPAK6-pgh was obtained successfully.SDS-PAGE and Western blot analysis using the infection cells showed that the band of 25 kD was particular to pGH gene product. The culture liquid of recombinant virus was injected into the individual of Bombyx mori, the special band of 25 kD rpGH still exist according to the SDS-PAGE and Western blot analysis.
出处
《蚕业科学》
CAS
CSCD
2004年第4期376-381,共6页
ACTA SERICOLOGICA SINICA
基金
江苏省教育厅自然科学基金项目 (编号Q1114 0 2 0 )
