[Expression of interferon alpha family gene of Chinese marmot in eukaryotic and prokaryotic cells].
OBJECTIVE
To investigate the function of interferon alpha (IFNalpha) in a Chinese marmot model of hepatitis B, we expressed the Chinese marmot (Marmota himalayana) IFNalpha family gene (IFNA) in eukaryotic cells and prokaryotic cells.


METHODS
Eukaryotic and prokaryotic expression plasmids harboring Chinese marmot interferon alpha gene with different genotypes were generated using molecular cloning technology. We detected the biological activity of all expression products by viral protection assay, and analyzed their differences and species restriction of the biological activity.


RESULTS
The Chinese marmot functional genotype IFNalpha was expressed in the baby hamster kidney (BHK) cell line, and these products protected WH12/6 cells challenged by encephalomyocarditis virus (EMCV). The Chinese marmot IFN-alpha5 also expressed in E. Coli induced by IPTG, and purified fusion protein had antiviral biological activity. The biologic activity displayed differences among different subtype IFNalpha, and it had strict species restriction.


CONCLUSIONS
The IFNalpha family gene of the Chinese marmot can be expressed in both eukaryotic and prokaryotic cells, and the expression products show antiviral activity in a protection assay. This study provides, for the first time, evidence that IFNalpha from the Chinese marmot has an antiviral function in vitro and can be used to improve the efficacy of current therapies for HBV infection in our Chinese marmot model.
Lu
Yin-ping
YP
Division of Clinical Immunology, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Wang
Bao-ju
BJ
Huang
Hong-ping
HP
Tian
Yong-jun
YJ
Yang
Yan
Y
Dong
Ji-hua
JH
Lu
Meng-ji
MJ
Yang
Dong-liang
DL
D000818:Animals; D005057:Eukaryotic Cells / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006509:Hepatitis B / Q000378:metabolism*; D016898:Interferon-alpha / Q000096:biosynthesis* / Q000235:genetics* / Q000502:physiology; D008392:Marmota / Q000378:metabolism*; D011387:Prokaryotic Cells / Q000378:metabolism; D015398:Signal Transduction; D014157:Transcription Factors
D016898:Interferon-alpha; D014157:Transcription Factors