Subclone of partial sequence of gene of HBV X region and PCR detection

Objective: To explore the significance and value of HBV X region genes in the early diagnosis of Hepatitis B.Methods: The partial gene sequence of HBV X region in pBR322-HBV was amplified by PCR.The products amplified by PCR were sub-cloned into the pBS-T vector,which were an effective AT clone plasmid.The positive AT sub-clones were sequenced.Results: The expected plasmid with HBV X region gene was obtained.The optimal annealing temperature was 51 ℃.The sensitivity reached 101 copies per 2μl.The linear range was 101-1010 copies per 2μl.Discussion: The target genes in pBR322-HBV were sub-cloned to pBS-T.The amplification product of the target gene sequence in pBR322-HBV was 57bp.It can be directly sub-cloned to the new vector without purification,which favors the routine PCR and TaqMan MGB fluorescent quantitative PCR for the early detection of HBV.