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- W2379099755 abstract "In ordder to screening the BHK-21cell lines stable expression of ST3GalⅠgene for further study on the AIV cell culture,the chicken ST3GalⅠgene was coloned and incorporated into the eukaryotic expression vector pcDNA3.1-EGFP to generate the recombinant plasmid pcDNA3.1-EGFP-ST3GaⅠ.The recombinant vector pcDNA3.1-EGFP-ST3GalⅠ was identified and transfected into BHK-21cells using lipofectamine 2000.ST3GalⅠ gene was stably expressed in cells.The monoclone cell line was obtained by G418resistance screening and RT-PCR.To detect proliferation of avian influenza virus(AIV)H9subtype and H5N1Re-5in BHK-21-ST3GalⅠcell,the different proportion of the H9subtype and H5N1Re-5avian influenza viruses were inoculated in BHK-21-ST3GalⅠ cells and blank BHK-21cells.At the same time,the ordinary pancreatic enzyme group and TPCK pancreatic enzyme control group were set up.The cell liquid was harvested after 72hours.The hemagglutinin(HA)titer was determined by conventional method.After being transfected into BHK-21cells,ST3GalⅠ was stably expressed in BHK-21.Green fluorescence could be observed and a 1 029bp cDNA fragment of expressed RNA was detected by RT-PCR.The experimental results showed that HA titer reached 1∶256in BHK-21-ST3GalⅠcells at the inoculation amount 2%,which is significantly higher than that in blank BHK-21cell group,and more suitable for the flu virus proliferation,having the potential for the influenza virus vaccine production." @default.
- W2379099755 created "2016-06-24" @default.
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- W2379099755 date "2013-01-01" @default.
- W2379099755 modified "2023-09-23" @default.
- W2379099755 title "Establishment of BHK-21 Cell Line Stably Expressing Chicken ST3GAL I Gene" @default.
- W2379099755 hasPublicationYear "2013" @default.
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