利用杆状病毒表达系统表达金鱼生长激素I基因

Expression of Goldfish Growth Hormone I Gene Using Baculovirus Expression System

以不含起始密码ＡＴＧ的质粒ｐＳＸＩＶＶＩ＋Ｘ３为转移载体，将编码金鱼生长激素Ｉ的ｃＤＮＡ插入粉纹夜蛾核型多角体病毒（ＴｎＮＰＶ）基因组中，构建了形成多角体的重组病毒株ＴｎＮＰＶＳＸ＋ｇｆＧＨＩ２ｌａ。该毒株能利用合成多角体ＸＩＶ串联启动子，在重组病毒感染的草地贪夜蛾（Ｓｐｏｄｏｐｔｅｒａｆｒｕｇｉｐｅｒｄａ，Ｓｆ）昆虫细胞及银纹夜蛾幼虫中表达金鱼生长激素Ｉ基因。感染离体细胞及虫体后的蛋白ＳＤＳ聚丙烯酰胺凝胶电泳表明，所表达的蛋白分子量为２２５ｋＤａ，与理论计算值相符。Ｗｅｓｔｅｒｎ印迹证实。

Using pSXIVVI +X3 without an initiation codon as an expressing vector, an occluded recombinant Trichoplusia ni nuclear polyhedrosis virus carrying the cDNA encoding goldfish growth hormone I (gf GHI) under the control of the Syn XIV promoter, has been constructed. Immunoblot analysis showed that the molecular weight of the expressed protein was 22 5kDa as expected, revealed that the virus mediated gfGHI was synthesized in the Sf cells, larvae and secreted into the supernatant and hemolymph.