| Hepatitis B virus (HBV) is the prototypic member of the hepadnavirus family, which causes acute and chronic liver diseases, including chronic active hepatitis, as well as cirrhosis and hepatocellular carcinoma. HBV infects an estimated 200 million persons worldwide and thus represents a viral pandemic. X gene is the smallest open reading frame of HBV. X gene codes for a 16.5KD protein (X protein, HBx), which is made of 154 amino acids. HBx is a multifunctional viral regulator that modulates transcription, cell responses to genotoxic stress, protein degradation, and signaling pathways. Besides, it may affect DNA repair, cell cycle control, and apoptosis. The HBx protein is essential for viral infectivity and is thought to play a role in development of hepatocellular carcinoma during chronic hepatitis B virus infection.To explore the possible function of HBV XTP12 after HBV infection, The recombined expression plasmid pcDNA3.1(-)- XTP12 was constructed, and HepG2 cells were transfected, and pcDNA3.1(-) empty vector was used as control. The mRNA was isolated from HepG2 cells transfected with pcDNA3.1(-)-XTP12 and pcDNA3.1(-) empty vector, respectively, and suppression subtractive hybridization (SSH) method was employed to analyze the differentially expressed DNA sequence between the two groups. The obtained production was subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E. coli strain DH5a in random. The cDNA was sequenced and analyzed in GenBank with Blast search after PCR.. At the same time, Microarray was employed for detecting and analyzing of both mRNA from the HepG2 cells. The obtained sequences may be target genes transactivated by XTP12, among which some genes coding proteins involved in cell signal transduction, cycle regulation, translation and synthesis of protein, metabolism) immunoregulation, and correlated with tumor.These may explain the possible mechanism of XTP12 in vivo. |
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