中国人兽共患病学报2012，Vol.28Issue(7)：674-678,5.DOI:10.3969/cjz.j.issn.1002-2694.2012.07.006
猴B病毒gB基因在昆虫细胞中的表达
Expression on gB gene of simian B virus in insect cells
摘要
Abstract
In order to obtain antigenic properties that used to construct a ELISA kit to test BV antibody and take the place of the BV, gB gene of simian B virus/E2490 was synthesized according to the reported BV virus gB gene (GenBank accession number; AF533768). The gB gene fragment was than cloned in the vector pFastBac-HTa. The recombinant plasmid pFastBac-HTa-gB was constructed and identified by PCR and enzyme digestion and sequenced. Then the plasmid pFast-HTa-gB was transformed into DHlOBac complement cells. It was identified by antibiotics and PCR, indicating that we have constructed baculovirus expression vector to gB gene. On this basis, transposition bacmid DNA was extracted to transfect sf9 insect cells. After transfected for 96 h, the sf9 insect cells showed expansion, round and cell nucleus turned to great, and baculovirus was harvested. Then the SDS-PAGE and Western blot and indirect immunofluorescence assay results showed the expression of gB protein in insect cells. The gB protein has a good biological activity and was approximate 98kDa. It was a base to construct ELISA kit to test BV antibody. The result showed that we have constructed the recombinant plasmid pFastBac-HTa-gB and baculovirus expression vector to gB gene, and the gB gene was expressed in insect cells.关键词
猴B病毒/gB基因/杆状病毒表达载体/昆虫细胞Key words
simian B virus (BV)/ gB Gene/ Baculovirus expression vector/ insect cells分类
医药卫生引用本文复制引用
段博芳,艾军,王琼,段纲,毛永杨,叶玲玲,杨建明,徐维加,董俊,周晓黎..猴B病毒gB基因在昆虫细胞中的表达[J].中国人兽共患病学报,2012,28(7):674-678,5.基金项目
国家质检总局科技计划项目(20091K009)资助,云南省高端科技人才进项目(2009C1125)资助 （20091K009）
