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KMID : 0380220070400061002
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Journal of Biochemistry and Molecular Biology
2007 Volume.40 No. 6 p.1002 ~ p.1008
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Overexpression and Purification of PreS Region of Hepatitis B Virus Antigenic Surface Protein adr Subtype in Escherichia coli
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Abbas Naaz
Ahmad Aftab
Shakoori Abdul Rauf
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Abstract
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PreS domain of Hepatitis B virus (HBV) surface antigen is a good candidate for an effective vaccine as it activates both B and T cells besides binding to hepatocytes. This report deals with overexpression and purification of adr subtype of surface antigen that is more prevalent in Pakistan. PreS region, comprising 119 aa preS1 region plus a 55 aa preS2 region plus 11 aa from the N-terminal S region, was inserted in pET21a+ vector, cloned in E. coli DH5α cells and expressed in E. coli BL21 codon+ cells. The conditions for over expression were optimized using different concentrations of IPTG (0.01-5 mM), and incubating the cells at different temperatures (23-41°C) for different durations (0-6 h). The cells were grown under the given optimized conditions (0.5 mM IPTG concentration at 37°C for 4 h), lysed by sonication and the protein was purified by ion exchange chromatography. On the average, 24.5 mg of recombinant protein was purified per liter of culture. The purified protein was later lyophilized and stored at -80°C.
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KEYWORD
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Adr subtype of surface antigen, Hepatitis B surface antigen, Hepatitis B virus, Hepatitis in Pakistan, PreS
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