Construction of the eukaryotic expression plasmid for mp65 gene of Candida albicans.

To construct the eukaryotic expression plasmid of the mp65 gene in Candida albicans and to analyze its immunogenicity,this gene was firstly amplified by using the PCR technique from the standard strain ATCC 64550 mice,and then inserted to eukaryotic expression vector pcDNA3.1.The eukaryotic expression plasmid was extracted and was used to immunize mice,while the antibodies produced against MP65 protein as well as the cell-mediated immune responses were detected by ELISA and flow cytometry respectively.In this way,the mp65 gene with a molecular size of 1140 bps was harvested by PCR and the eukaryotic expression plasmid pcDNA3.1-mp65 was constructed successfully.This recombinant plasmid used to immunize BALB/c mice could induce the production of IgG antibodies with a serum titer of 1:800.Also,the percentages of CD4+ and CD8+ T lymphocytes were elevated.It is evident that the humoral and cell-mediated immune responses can be induced by the immunization of mice with the eukaryotic expression plasmid pcDNA3.1-mp65.