水产科技情报2025，Vol.52Issue(1)：26-32,7.DOI:10.16446/j.fsti.20220700107
大西洋鲑传染性胰腺坏死病病毒VP2蛋白和VP3蛋白重组腺病毒载体的构建及其表达
Construction and expression of recombinant adenovirus vector against infectious pancreatic necrosis with VP2 protein and VP3 protein of Atlantic salmon
摘要
Abstract
Infectious pancreatic necrosis(IPN)was an infectious disease caused by infectious pancreatic necrosis virus(IPNV),which mainly affected salmonids.To clone the infectious pancreatic necrosis virus(IPNV)VP2 gene and VP3 gene,constructed their recombinant adenovirus vector.In this study,the IPNV VP2 gene and VP3 gene were amplified and inserted into the pAd-Track-CMV vector by PCR and multi-gene fragment homologous re-combination technology.After linearization,the recombinant plasmids were recombined with the pAd-easy-1 vec-tor in E.coli BJ5183 to construct a recombinant adenovirus plasmid,which was identified by PCR and digestion with NotⅠand HindⅢ,and then transfected into 293T cells to obtain the recombinant adenovirus after linearizing with PacⅠ.The viral replication was monitored using the expression of green fluorescent protein(GFP).The ex-pression of VP2 protein and VP3 protein were confirmed by the Western-blotting method,and the recombinant viral titer was detected.The results indicated that the IPNV VP2 gene and VP3 gene were cloned with a length of 2 211 bp,and the recombinant adenovirus vectors for the target genes were constructed.The virus could express a molecular mass of about 54 kD and 26 kD in 293T cells,respectively,with a titer of 1.0×109 cell/mL TCID50.The findings revealed that the recombinant adenovirus with the IPNV VP2 gene and VP3 gene was successfully ob-tained,and the virus had a high titer in 293T cells and could be expressed stably.The successful construction of recombinant adenovirus vectors with VP2 and VP3 proteins of IPNV could provide a reference for the development of the vaccine and better prevention of IPNV in Atlantic salmon.关键词
大西洋鲑/传染性胰腺坏死病/VP2基因/VP3基因/腺病毒载体Key words
Atlantic salmon/infectious pancreatic necrosis/VP2 gene/VP3 gene/adenovirus vector引用本文复制引用
李守湖,秦闯,李新苍,石建高..大西洋鲑传染性胰腺坏死病病毒VP2蛋白和VP3蛋白重组腺病毒载体的构建及其表达[J].水产科技情报,2025,52(1):26-32,7.基金项目
上海市启明星项目——扬帆专项(22YF1459800) （22YF1459800）
中央公益性科研院所基本业务费(2024XT0506,2020TD43). （2024XT0506,2020TD43）
