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Expression in Escherichia coli of a cloned DNA sequence encoding the pre-S2 region of hepatitis B virus

Offensperger, Wolf and Wahl, Silke and Neurath, A. Robert and Price, Peter and Strick, Nathan and Kent, Stephen B. H. and Christman, Judith K. and Acs, George (1985) Expression in Escherichia coli of a cloned DNA sequence encoding the pre-S2 region of hepatitis B virus. Proceedings of the National Academy of Sciences of the United States of America, 82 (22). pp. 7540-7544. ISSN 0027-8424. PMCID PMC390852. doi:10.1073/pnas.82.22.7540.

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A DNA sequence encoding the entire pre-S2 region (amino acids 120-174; serotype ayw) of human hepatitis B virus envelope protein has been inserted into the lacZ gene of the plasmid pSKS105 yielding a recombinant, pWS3. Lac+ colonies of the Escherichia coli M182 (lacIOPZYA), isolated after transformation with pWS3, produced a pre-S2 peptide-ß-galactosidase fusion protein. This fusion protein, which comprised as much as 3% of the total bacterial protein, was purified to >90% homogeneity by affinity chromatography on p-aminophenyl-ß-D-thiogalactoside-Sepharose. It is immunoprecipitable with rabbit antibodies to a synthetic peptide corresponding to amino acids 120-145 of the pre-S2 region of serotype adw [pre-S(120-145)] or with antibodies to hepatitis B virus. pre-S(120-145) completely blocked the binding of either antibody to the pre-S2 peptide-ß-galactosidase fusion protein. These results indicate that there are antigenic determinants on the fusion protein that are closely related to, if not identical to, determinants on synthetic pre-S(120-145) and on pre-S2 sequences of native hepatitis B virus particles. Thus, bacteria transformed with pWS3 can provide an abundant source of pre-S2-ß-galactosidase fusion protein, which may prove useful either as a diagnostic reagent possessing marker enzyme activity suitable for ELISA tests or as an immunogen with potential to contribute to active prophylaxis of hepatitis B.

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Additional Information:© 1985 by the National Academy of Sciences. Communicated by Hans Popper, July 18, 1985. W.O. and S.W. are Fellows of the Deutsche Forschungsgemeinschaft. This investigation was partially supported by Public Health Service Grant RO1-CA34818 awarded by the National Cancer Institute. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Funding AgencyGrant Number
Deutsche Forschungsgemeinschaft (DFG)UNSPECIFIED
Subject Keywords:recombinant DNA; ß-galactosidase fusion protein; affinity chromatography; antigenic determinants of the hepatitis B virus envelope
Issue or Number:22
PubMed Central ID:PMC390852
Record Number:CaltechAUTHORS:OFFpnas85
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:9235
Deposited By: Tony Diaz
Deposited On:28 Nov 2007
Last Modified:08 Nov 2021 20:57

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