[Expression of influenza A3 virus (H3N2) M2 gene in vaccinia virus Tiantan strain].
To construct a recombinant vaccinia virus RVJ1175M2 expressing influenza A3 virus (H3N2) M2 gene, full length gene encoding influenza virus (H3N2) M2 protein was amplified with PCR and cloned into plasmid pJSC1175 which was used for homologous recombination with vaccinia virus Tiantan strain. Along with this, a recombinant vaccinia virus RVJ1175M2 containing the M2 gene was subsequently constructed. It was identified by PCR that the gene of M2 protein was inserted into the TK locus of vaccinia virus Tiantan strain correctly and M2 protein was expressed by recombinant vaccinia virus RVJ1175M2 effectively. Two electrophoretic bands of M2 protein expressed by the infected HeLa cells, one of 15kD and the other of 13kD in accordance with related documents, was deteced by Western-blot. M2 protein distributing on the surface of the infected cells was demonstrated by immunofluorescence and flow cytometry. The results suggested that recombinant vaccinia virus RVJ1175M2 could express M2 protein effectively, this laid a foundation for comparative research on the immune effect of universal vaccine of influenza virus with different kinds of vaccine expressing M2 protein.
Wang
Wen-Ling
WL
Biotech Center for Viral Disease Emergency, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China. wangwenling_ivdc@yahoo.com.cn
Huang
Bao-Ying
BY
Deng
Yao
Y
Wang
Xiu-Ping
XP
Tan
Wen-Jie
WJ
Ruan
Li
L
D006367:HeLa Cells; D006801:Humans; D053122:Influenza A Virus, H3N2 Subtype / Q000235:genetics*; D007252:Influenza Vaccines / Q000276:immunology; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis; D014614:Vaccines, Synthetic / Q000276:immunology; D014616:Vaccinia virus / Q000235:genetics*; D014763:Viral Matrix Proteins / Q000235:genetics*; D000086082:Viroporin Proteins
D007252:Influenza Vaccines; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D014763:Viral Matrix Proteins; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins