应用重组家蚕核多角体病毒在蚕体中表达人丙型肝炎病毒C区及E1区基因

Expression of C and E1 Gene of HCV in Silkworm With Recombinant BmNPV

将人丙型肝炎病毒的C区、E1区及部分E2区基因装入转移载体 pBM0 3 0 ,经原位杂交筛选、酶切鉴定、Southern杂交检测确定转移质粒装载成功。将转移质粒与家蚕核多角体病毒重组 ,在家蚕BmN细胞中挑选重组病毒 ,经点杂交技术确证重组病毒带有目的基因。将重组家蚕核多角体病毒通过针刺和注射方法感染蚕蛹及 5龄期家蚕 ,肽抑制试验检测目的基因表达的多肽活性。结果表明 ,目的基因已得到表达 ,表达量较高 ,为每蛹体约 60～ 60 0 μg。

A plasmid transfer vector containing the C,E1 and a part of E2 genes of human HCV was constructed through inserting these genes into the plasmid pBM030.Then the transfer vecter DNA was introduced into BmN cells along with wild type genomic visal DNA of Bombyx mori nuclear polyhedrosis virus (BmNPV).The recombinant BmNPV containing the foreign genes was identified by plaque assays and by dot bloting.It is infectious to larva and pupa of the silkworm and can express the polypeptide encoded by the foreign genes in the silkworm at a high level of 60～600μg per pupa.The biological activity of the expressed polypeptide was detected by polypeptide inhibiting test.