Research on HBV Gene Integration into Host Genome that is Related to HBV DR Region
Department of medicine, Southwest of Guizhou Vocational and Technical College for Nationalities, Xingyi 562400, China
Genomics and Applied Biology, 2016, Vol. 7, No. 5 doi: 10.5376/gab.2016.07.0005
Received: 04 Jul., 2016 Accepted: 06 Sep., 2016 Published: 09 Nov., 2016
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Preferred citation for this article:
Luo H.X., Gu L., He L.H., 2016, Research on HBV Gene Integration into Host Genome that is related to HBV DR region, Genomics and Applied Biology, 7(6): 1-8 (doi: 10.5376/gab.2016.07.0006)
In most studies, the HBV DNA has been found integrated into the DNA of the hepatocellular carcinoma cells, the integrated HBV DNA is related to HBV DR region, in order to research how HBV integrated into the genome of the cells via HBV DR region, we isolated HBV Creg DNA fragment which is from nt1087 to nt2488 contained the DR region, regulatory sequence, X gene and C gene and constructed pcDNA3.1(+)-HBV Creg eukarya expression vector, then transfected it into the HepG2 cells to observe the integration way of HBV. In this study, HBV Creg DNA fragment was isolated from HBV genome by recombinant PCR, in this DNA fragment, the DR region, regulatory sequence, X gene and C gene of HBV was included, then linked it to the vector pcDNA3.1(+) to construct the pcDNA3.1(+)-HBV Creg eukarya expression vector. We transfected the pcDNA3.1(+)-HBV Creg eukarya expression vector into HepG2 cells, in the process, the transfected cell lines were selected by G418, the total DNA of HepG2 cells was extracted to test the integration of HBV by PCR using different primers. The full length of HBV Creg DNA fragment contained DR region could only be detected by PCR before integration into cell genome, after the fragment integrated into HepG2 genome, the full length of HBV Creg DNA fragment had not been detected all the time, however, HBV X gene and HBV C gene could be detected respectively, for the reason of HBV DR region, it may integrate at DR1 or DR2 with the Creg DNA extending downstream or upstream though C gene or X gene. The HBV Creg fragment can integrate into HepG2 cells genome. And the integration of HBV is related to DR region, the way of HBV Creg DNA fragment integrated into HepG2 cells is from DR region to both sides.
Hepatitis B virus; Hepatocellular carcinoma; Gene Recombinant; DR region; Gene integration
Genomics and Applied Biology
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