[Construction of eukaryotic expression vector expressing hepatitis B virus HBsAg and EGFP fusion protein and establishment of stable transfected Chang Liver cell line].
OBJECTIVE
To construct a eukaryotic expression vector for expressing hepatitis B virus (HBV) recombinant HBsAg-EGFP fusion protein and obtain a stable transfected Chang Liver cell line.


METHODS
The coding region of HBsAg gene of HBV was amplified by PCR and was digested by BamH I/EcoR I . This fragment was inserted into pEGFPN1 with T4 ligase and transformed E-coli TG1. The positive recombinant plasmid was selected, then the recombinant plasmid was transfected into Chang Liver cell by Lipofectamine 2000 cells containing stable transformants were selected by the ability of resistance to G418 and isolated with a limited dilution. The stable transfected cell line expressing high level HBsAg-EGFP fusion protein was obtained.


RESULTS
The eukaryotic expression vector named pEGFPN1-HBsAg was successfully constructed and the stable transfected Chang Liver cell line expressing pEGFPN1-HBsAg fusion protein was obtained.


CONCLUSIONS
The stable transfected Chang Liver cell line could express pEGFPN1-HBsAg fusion protein, could be used to screen the proteins differentially expressed in HBsAg expression Chang Liver cells, which brought some new clues for studying the potential molecular mechanism of HBsAg protein.
Mu
Jin-Song
JS
PLA Postgraduate Medical School, Beijing 100039, China.
Wang
Hui-Fen
HF
Wang
Jiang-Hua
JH
Pan
Xiao-Ben
XB
Zhang
Lu
L
Wei
Lai
L
D002460:Cell Line; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D006514:Hepatitis B Surface Antigens / Q000235:genetics* / Q000378:metabolism; D006515:Hepatitis B virus / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection / Q000379:methods*
D006514:Hepatitis B Surface Antigens; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins