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

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 (Ni2+) chelating affinity chromatography. The immunogenicity was tested by Western blot. RESULTS: The length of the fragment gene was 243bp, coding 81 amino acids; the relative molecular mass of recombinant protein was 9000. 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.