Construction and expression of hepatitis B virus
vector encoding TC-tagged core protein
Construction and expression of hepatitis B virus
vector encoding TC-tagged core protein
Yuanyuan LIN MD1,Xiaoming CHENG MS1,Yuhu SONG MD1,Peiyuan LI MD1,Ying CHANG MM1,Jinjian YAO MD1,Jusheng LIN MD, PhD1,Li ZHOU PhD2,Leiming XU PhD3,
1.Institute of Liver Diseases,
Tongji Hospital, Tongji Medical College, Huazhong University of Science
and Technology, Wuhan 430030, China; 2.Department of Neurobiology,
Tongji Medical College, Huazhong University of Science and Technology,
Wuhan 430030, China; 3.Department of Pathophysiology,
Tongji Medical College, Huazhong University of Science and Technology,
Wuhan 430030, China;
Abstract：Virus tagged with green fluorescent protein (GFP) contributes to the visualization and study of the virus in living cells. However, the hepatitis B virus (HBV) particle, which is a compact virion with limited internal space, cannot be incorporated with GFP tag as a large fragment. It was recently reported that protein genetically inserted with a smaller size tetracysteine (TC) tag could be specially labeled by a biarsenical fluorescent dye in living cells. In this study, we constructed a recombinant HBV vector encoding TC-tagged core protein for biarsenical labeling of HBV virion. TC tag was genetically inserted near the immunodominant c/e1 site of HBV core protein by mutagenesis. Western blot and enzyme-linked immunosorbent assay (ELISA) analysis showed that the TC-tagged core protein, hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) could be expressed in cells transfected with the recombinant HBV vector, which is similar to the cells transfected with wild-type HBV vector. Reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot analysis showed that HBV virion formation was affected by the genetic insertion of TC tag into core protein in some degree, but cells transfected with the HBV vector could still produce HBV virions incorporated with TC-tagged core proteins. Taken together, the recombinant HBV vector can serve as a useful tool to produce HBV virions incorporated with TC-tagged core proteins to be fluorescently labeled by biarsencial dye for visualizing and studying HBV in living cells.
. Construction and expression of hepatitis B virus
vector encoding TC-tagged core protein[J]. Front. Med., 2009, 3(4): 396-402.
Yuanyuan LIN MD, Xiaoming CHENG MS, Yuhu SONG MD, Peiyuan LI MD, Ying CHANG MM, Jinjian YAO MD, Jusheng LIN MD, PhD, Li ZHOU PhD, Leiming XU PhD, . Construction and expression of hepatitis B virus
vector encoding TC-tagged core protein. Front. Med., 2009, 3(4): 396-402.
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