[Clone of hepatitis B virus X gene and its protein expression].
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.


CONCLUSIONS
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.
Tang
Shi-Gang
SG
Department of Infectious Diseases, Third Xiangya Hospital, Central South University, Changsha 410013, China. sjtangq@163.com
Yang
Bing-Chang
BC
D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D017422:Sequence Analysis, DNA; D015534:Trans-Activators / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection; D054334:Viral Regulatory and Accessory Proteins
D011994:Recombinant Proteins; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein