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Journal of Central South University(Medical Sciences) ; (12): 525-528, 2005.
Article in Chinese | WPRIM | ID: wpr-813514

ABSTRACT

OBJECTIVE@#To construct the hepatitis B virus (HBV) X gene recombinant and to induce the expression of X protein.@*METHODS@#HBV DNA was extracted from the serum of patient with hepatitis B. The X gene was amplified by PCR using the primers with EcoRI and HindIII digestion sites, and then cloned into pronucleus expression vector pMAL-C2X, which was detected by EcoRI and HindIII digestion and sequence. Finally, the recombinant was induced by IPTG to express X protein in JM109.@*RESULTS@#The band similar to X gene was amplified by PCR. There were fragments similar to X gene when the recombinant was digested by the enzyme digestion. It was tested by DNA sequence that the correct and entire opening reading frame of HBV X gene was inserted. The X protein was expressed by the IPTG induction.@*CONCLUSION@#Pronucleus expression recombinant pMAL-C2X-HBV-X is constructed successfully and with the IPTG induction, the recombinant pMAL-C2X-HBV-X can express the X protein in E. coli JM109, which lays the foundation for the HBV X protein purification and its biological study.


Subject(s)
Female , Humans , Male , Base Sequence , Cloning, Molecular , Escherichia coli , Genetics , Hepatitis B virus , Genetics , Molecular Sequence Data , Open Reading Frames , Genetics , Recombinant Proteins , Genetics , Sequence Analysis, DNA , Trans-Activators , Genetics , Transfection
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