[Transient expression of fusion protein with a chimeric HBsAg-HSP70 construct in HepG2 cells].
OBJECTIVE
To investigate transient expression of fusion protein with a chimeric HBsAg-HSP70 construct in HepG2 cells.


METHODS
Enkaryotic expression plasmids inserted HBsAg gene or chimeric HBsAg-HSP70 gene were prepared and transfected into HepG2 cells by means of cationic liposome. mRNA were detected by RT-PCR and proteins expressed in the cells were detected by immunocytochemistry 48 hours later. HBsAg in cultured supernatants and cell lysates were assayed by ELISA.


RESULTS
Fusion protein (HBsAg-HSP70) transient expression in HepG2 cells were confirmed by RT-PCR, immunocytochemistry or ELISA, but fusion protein was not assayed in cell cultured supernatants by ELISA.


CONCLUSIONS
Transfection of HepG2 cells with a chimeric HBsAg-HSP70 construct leads to express fusion protein, but it does not secrete into cell cultured supernatants.
Liang
Zeng Wei
ZW
Institute for Viral Hepatitis, Chongqing University of Medical Science, Chongqing 400010, China.
Lan
Ying Hua
YH
Ren
Hong
H
Li
Yong Guo
YG
Cai
Da Chuan
DC
D001426:Bacterial Proteins; D004797:Enzyme-Linked Immunosorbent Assay; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics*; D006514:Hepatitis B Surface Antigens / Q000235:genetics*; D017325:Hepatitis B Vaccines / Q000276:immunology; D006801:Humans; D007150:Immunohistochemistry; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019444:Vaccines, DNA / Q000276:immunology
D001426:Bacterial Proteins; D018840:HSP70 Heat-Shock Proteins; C444642:HSP70 protein, Mycobacterium tuberculosis; D006514:Hepatitis B Surface Antigens; D017325:Hepatitis B Vaccines; D011993:Recombinant Fusion Proteins; D019444:Vaccines, DNA