Secreted Expression of Human Beta-defensin-3 in Saccharomyces cerevisiae
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Abstract:
DNA fragment containing hBD-3 with α-factor in the upstream sequence was amplified and inserted into the shuttle plasmid-pYES2 of Saccharomyces cerevisiae after digestion. The recombinant vector called pYES2-α-factor-hBD3 was transformated into Saccharomyces cerevisiae INVSC1. The recombinant positive clones, which were screening by PCR reduced with 2% galactose. The recombinant protein could be detected in the cultured supernatant by protein assay reagents. Plate test indicated that the supematant could inhibit the growth of Escherichia coli ATCC6538 and Staphylococcus aureus ATCC10231. The strategy of secretion expression of Human beta-defensin-3 gene in Saccharomyces cerevisiae may lay the foundation for further exploration of the biological activity and security application of the Human beta-defensin-3.
