Production of hepatitis B virus preS polypeptide in Escherichia coli by mutation of the 5'-end coding sequence and its purification and characterization

Hee Sun Kim, Youn Kyu Kim, Seong Eon Ryu, Hyo Jeong Hong

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The preS1 and preS2 antigens (preS Ag) of hepatitis B virus (HBV) elicit virus-neutralizing and protective antibodies which can overcome nonresponsiveness to the currently available vaccine for HBV and also carry the attachment site to HBV hepatocyte receptor. Therefore, in order to study the development of more effective vaccine and the receptor-ligand interaction, it will be helpful to obtain high-level production of the preS Ag from bacteria. We have found that the native preS region gene was not expressed under the control of commonly used promoters in Escherichia coli. By site-directed mutagenesis of some nucleotides at the 5'-end of the preS1 region gene, we have generated a mutant gene which is highly expressed in soluble form in E. coli. The produced polypeptide could be efficiently purified by 20% ammonium sulfate precipitation and a gel permeation chromatography and the purified polypeptide was demonstrated to exhibit the antigenicity and the immunogenicity of the preS1 and preS2 Ag, suggesting that it is functional.

Original languageEnglish
Pages (from-to)173-177
Number of pages5
JournalGene
Volume177
Issue number1-2
DOIs
StatePublished - 1996 Oct 24

Fingerprint

Hepatitis B virus
Escherichia coli
Peptides
Mutation
Vaccines
Genes
Virus Receptors
Antigens
Ammonium Sulfate
Site-Directed Mutagenesis
Neutralizing Antibodies
Gel Chromatography
Hepatocytes
Nucleotides
Ligands
Viruses
Bacteria

Keywords

  • Antigenicity
  • Expression
  • Hepatitis B virus
  • Immunogenicity
  • Mutation
  • Purification
  • Recombinant DNA
  • preS

Cite this

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title = "Production of hepatitis B virus preS polypeptide in Escherichia coli by mutation of the 5'-end coding sequence and its purification and characterization",
abstract = "The preS1 and preS2 antigens (preS Ag) of hepatitis B virus (HBV) elicit virus-neutralizing and protective antibodies which can overcome nonresponsiveness to the currently available vaccine for HBV and also carry the attachment site to HBV hepatocyte receptor. Therefore, in order to study the development of more effective vaccine and the receptor-ligand interaction, it will be helpful to obtain high-level production of the preS Ag from bacteria. We have found that the native preS region gene was not expressed under the control of commonly used promoters in Escherichia coli. By site-directed mutagenesis of some nucleotides at the 5'-end of the preS1 region gene, we have generated a mutant gene which is highly expressed in soluble form in E. coli. The produced polypeptide could be efficiently purified by 20{\%} ammonium sulfate precipitation and a gel permeation chromatography and the purified polypeptide was demonstrated to exhibit the antigenicity and the immunogenicity of the preS1 and preS2 Ag, suggesting that it is functional.",
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Production of hepatitis B virus preS polypeptide in Escherichia coli by mutation of the 5'-end coding sequence and its purification and characterization. / Kim, Hee Sun; Kim, Youn Kyu; Ryu, Seong Eon; Hong, Hyo Jeong.

In: Gene, Vol. 177, No. 1-2, 24.10.1996, p. 173-177.

Research output: Contribution to journalArticle

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AU - Kim, Youn Kyu

AU - Ryu, Seong Eon

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AB - The preS1 and preS2 antigens (preS Ag) of hepatitis B virus (HBV) elicit virus-neutralizing and protective antibodies which can overcome nonresponsiveness to the currently available vaccine for HBV and also carry the attachment site to HBV hepatocyte receptor. Therefore, in order to study the development of more effective vaccine and the receptor-ligand interaction, it will be helpful to obtain high-level production of the preS Ag from bacteria. We have found that the native preS region gene was not expressed under the control of commonly used promoters in Escherichia coli. By site-directed mutagenesis of some nucleotides at the 5'-end of the preS1 region gene, we have generated a mutant gene which is highly expressed in soluble form in E. coli. The produced polypeptide could be efficiently purified by 20% ammonium sulfate precipitation and a gel permeation chromatography and the purified polypeptide was demonstrated to exhibit the antigenicity and the immunogenicity of the preS1 and preS2 Ag, suggesting that it is functional.

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