Abstract

Objective To clone the coding sequence of Staphylococcus aureus enterotoxin A (SEA) and to analyze its antigenicity for attenuation in targeted immunotherapy of tumor.Methods Genomic DNA extracted from Staphylococcus aureus 26079 was used as template.The coding special region of SEA was used as primer. Touchdown PCR method was employed to clone the coding region of mature SEA. PCR product recovered was subcloned into the pGEM-T vector. Positive clones were validated and selected by colony PCR followed by further confirmation by DNA sequencing and BL2SEQ alignment. The antigenicity of the coded peptide was analyzed by bioinformatic software.Results Following PCR product recovery and cloning,several clones were positively confirmed by colony PCR. DNA sequence alignment showed that the clones were of 100% identity with that of SEA registered in GenBank. Online analysis by SYFPEITHI showed that relatively high antigenicity was harbored in the whole region of mature SEA peptide with some extremely high peak values scattered in different regions.Conclusions The gene fragment encoding SEA mature peptide is successfully cloned. It is necessary to confirm all the loci with high peak antigenicity in order to attenuate the antigenicity of the peptide. Key words: Staphylococcus aureus; Enterotoxins; Genecloning; Antigenic variation

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