Construction of eukaryotic vector for HN gene of NDV Italien strain and its expression
- VernacularTitle:天然溶瘤新城疫病毒Italien株HN基因真核表达载体的构建和表达
- Author:
Hua ZHANG
;
Huijie BIAN
;
Ding WEI
- Publication Type:Journal Article
- Keywords:
newcastle disease virus;
hemagglutinin-neuraminidase gene;
transfection;
gene expression
- From:
Medical Journal of Chinese People's Liberation Army
1983;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct an eukaryotic expression plasmid containing gene coding for the hemagglutinin-neuraminidase(HN)of newcastle disease virus(NDV)oncolytic strain Italien,and then to express the protein in eukaryotic cell.Method The HN cDNA was synthesized from viral RNA by RT-PCR,and the eukaryotic expression vector of HN gene(named pcDNA3.1-HN)was constructed.The vector pcDNA3.1-HN was transfected into CHO-K1 cell by liposome,and G418 was used to select stable clones expressing HN gene.The expression of HN protein was visualized by Western blot and Immunofluorescence microscopy.Results Restriction analysis and DNA sequencing proved that HN gene was correctly cloned into expression vector.Western blot analysis and immunofluorescence showed that the HN was expressed in CHO-K1 cells.Conclusion The HN cDNA of NDV was successfully cloned into eukaryotic vector which showed good expression of HN protein in CHO-K1 cells.
