[Cloning and expression of a fragment of Corylus heterophylla 1 and identifying its immunologic competence]

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2011 Jan;27(1):4-6.
[Article in Chinese]

Abstract

Aim: To clone, express, and identify a fragment of Cor h 1 from Corylus heterophylla.

Methods: Through bioinformatics predication, the antigenic epitope of Cor h 1 was selected. A fragment gene of Cor h 1 was amplified by PCR and cloned into pMD18-T vector for sequencing. Then the fragment gene was sub cloned into pET-32a expression vector for expression, and then purified by metal (Ni(2+);) chelating affinity chromatography. The immunogenicity was tested by Western blot.

Results: The length of the fragment gene was 243 bp, coding 81 amino acids; the relative molecular mass of recombinant protein was 9 000. And the fragment of Cor h 1 was mainly expressed as soluble protein, purified protein has good immunogenicity.

Conclusion: The fragment gene of Cor h 1 was successfully cloned and expressed in this study, and the recombinant protein possessed good IgE-binding capacity.

MeSH terms

  • Allergens / chemistry
  • Allergens / genetics*
  • Allergens / immunology*
  • Amino Acid Sequence
  • Chromatography, Affinity / methods
  • Cloning, Molecular / methods
  • Corylus / chemistry
  • Corylus / genetics*
  • Corylus / immunology*
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • Genetic Vectors / chemistry
  • Genetic Vectors / genetics
  • Immunoglobulin E / immunology
  • Molecular Sequence Data
  • Nut Hypersensitivity / immunology
  • Plant Proteins / chemistry
  • Plant Proteins / genetics*
  • Plant Proteins / immunology*
  • Plasmids / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Sequence Analysis, DNA / methods

Substances

  • Allergens
  • Plant Proteins
  • Recombinant Proteins
  • Immunoglobulin E