关键词: 同源重组, ORF19.6012, MYC标签

Abstract:

Objective To construct ORF19.6012-MYC strain in Candida albicans. Methods The recombinant fragment composed of the upstream, downstream of the ORF19.6012 and the MYC-SAT1 flipper was constructed by In-Fusion kit. Then the fragment was homologously recombined into the genomic DNA of SN152 by lithium acetate transfection. The MYC tag was added to the C terminus of the open reading frame of ORF19.6012. PCR and Western blotting were used to verify the positive strain. Finally, the growth curve and hyphal formation ability of the positive strain were determined. Results ORF19.6012-MYC fusion strain was successfully constructed. The growth and proliferation ability of ORF19.6012-MYC strain and the ability of hyphal formation were consistent with those of the parental strain. Conclusion The ORF19.6012-MYC strain was successfully constructed and would be used in the following experiments.

Key words: homologous recombination, ORF19.6012, MYC tag