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A modified PCR system for amplifying beta-ketoacyl-ACP synthase gene fragments with DNA from Streptomyces
Feng-Ming Yu1, Xin Jiang, Jin-Chuan Wu
1Department of Pharmaceutical Engineering, Tianjin University, P.O. Box 6888, 300072, Tianjin, PR China.
Abstract:
Streptomyces luteogriseus strain 099, producing a new type of macrolide antibiotic with anti-coxB6 virus and anti-HIV protease activities, was isolated from soil. PCR was optimized to amplify beta-ketoacyl-ACP synthase (KS) genes. The system was optimized around the use of higher concentrations of DMSO (15% vs. 10% v/v) and dNTP (500 microM vs. 50-200 microM) and a lower annealing temperature (55 degrees C vs. 60-70 degrees C) than the normal PCR method used to amplify high GC content DNA.
