Team:HKUST/Group2

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Gene deletion

We are using a PCR-based gene deletion strategy (Baudin et al., Nucl. Acids Res. 21, 3329-3330, 1993 and Wach et al., Yeast 10, 1793-1808, 1994). Basically, knock-out fragments are generated by PCR, transformed into yeast strain, and integrated into the yeast genome by homologous recombination. Successful deletion replaces the yeast ORF with antibiotic resistant gene present in the knock-out fragment which serves as the selection marker.

Cassette plasmids, PCR primers and PCR conditions for amplification of knock-out fragments are chosen or designed according to a gene deletion toolbox (C. Janke et al., Yeast 2004; 21: 947–962.)