The Construction Of Expressing In Fluenza Virus A's PB2 And PA Proteins In 293T And MDCK Cell Lines

Influenza A virus has repeatedly occurred in recent years,and there have been many large-scale influenza epidemics,such as the Spanish flu in 1918,the Asian flu in 1957,the Hong Kong flu in 1968,and the influenza A H1N1 in 2009.All influenza epidemics have caused great losse to human,and the frequent occurrence of influenza A posed a great threat on the public health and safety.Therefore,the presence of influenza A virus vaccine,plays a vital role for human beings,the vaccine is the most effective measure to prevent influenza.Current influenza vaccines include inactivated and attenuated live vaccines.Inactivated vaccines include whole virus inactivated vaccines,splitting inactivated vaccines and subunit vaccines.When the virus matches to the vaccine strain,the protection rate of the inactivated vaccine to the young and middle-aged population was 66-86%.Both inactivated and attenuated live vaccines are chicken-derived vaccines.When using the chicken embryo as a matrix to produce influenza vaccine,there are external pollution and potential risk that vaccination with chicken infectious diseases may spread to human.In addition,the cycle of supplying chicken embryo is very long.When a big flu arrives,it is difficult to carry out large-scale production,especially,chicken embryo sources are more limited.The use of cell lines to produce vaccines can improve the defects that of using chicken embryo for production.This study is to construct 293 T and MDCK cell lines that stably expressing influenza protein PB2 and PA,combining with influenza virus reverse genetics technology system,which will provide strong technical support for the development of more secure replication-defective influenza virus attenuated live vaccine and contribute to human health.First,2009 H1N1 Influenza Virus adapted to save the virus UI182 was selected as a virus strain.MDCK-PB2 and MDCK-PA cell lines stably transfected with influenza virus MDCK cells were constructed and the cell biological characteristics of the cell lines,including cell morphology,cell proliferation,cell apoptosis and clone formation ability,were detected,and compared with the normal MDCK cell morphology;293T-PB2 and 293T-PA cell lines stably transfected with influenza virus 293 T cells were constructed and the cell morphology of the cells was detected as the basis for the rescue of influenza virus.In this study,we established MDCK-PB2,MDCK-PA cell line and 293T-PB2 and 293T-PA cell lines stably expressing the influenza virus MDCK.After the cell biology test of the cell line,the cell line was stable,Grow.The cell line is expected to play a positive role in the replication of defective vaccines after influenza virus replication.