Expression of HBV Pre S1 peptide in E. coli and product characterization.
HBV Pre S1 sequence is supposed to play an important role in the infection of HBV. Presence of Pre S1/anti-Pre S1 in serum has valuable clinical implications. In order to improve the study of Pre S1, Pre S1 sequence was overexpressed in E. coli as a fusion protein with MBP (Maltose-binding protein), and anti-Pre S1 antiserum was elicited in rabbits by Pre S1-MBP purified by affinity chromatography. The recombinant plasmid constructed from pMAL-cRI expressed the 106aa Pre S1 sequence at the C terminal of MBP by tac promoter. The resulting protein is about 54 kD in size. Western-blot analysis confirmed its reactivity with antiserum derived from synthetic Pre S1 peptide and serum from patients with acute hepatitis B (AHB). ELISA showed that Pre S1-MBP and Dane particles purified from AHB patient's serum reacted with antiserum against synthetic Pre S1 peptide, and this reaction was specifically inhibited by synthetic Pre S1 peptide. ELISA also demonstrated that antiserum against Pre S1-MBP reacted with synthetic Pre S1 peptide, but not with synthetic HCV peptide or HEV peptide.
Institute of Microbiology and Epidemiology, Beijing 100071.
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Z
Mi
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J
Guo
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Feng
W
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Chen
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Zhang
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Tong
D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis*; D015514:Oncogene Proteins, Fusion; D011498:Protein Precursors / Q000096:biosynthesis*; D014759:Viral Envelope Proteins / Q000096:biosynthesis*
D006514:Hepatitis B Surface Antigens; D015514:Oncogene Proteins, Fusion; D011498:Protein Precursors; D014759:Viral Envelope Proteins; C050806:presurface protein 1, hepatitis B surface antigen