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coli containing plasmid pANT35 or pANT62 was inoculated from an overnight culture at a ratio of 1:100 in a 2 l baffled Erlenmeyer flask filled with 400 ml Zym-5052 medium 51 containing 50 g ml 1 kanamycin sulfate per flask, incubated at 20 C, 140 rpm, 5 cm shaking diameter for 24 h, and the final OD 600 was determined using a CO8000 cell density meter (WPA biowave)