Application of RAPD Assays in Analyzing Streptomyces Gilvosporeus Strains from Genome Shuffling

摘要

In this study, the RAPD (Random Amplified Polymorphic DNA) technology was used to identify the genotypic variation between the parental strain and the shuffled strain of Streptomyces gilvosporeus. Firstly, RAPD conditions were optimized by orthogonal test. The results were as follows: DNA 60-150 ng, Taq DNA polymerase 1.0-1.5U, primer concentration 0.3-0.4 mmol/L, dNTPs concentration 200-250 µmol/L, Mg2+ concentration 2.5-3.0 mmol/L in 20µL PCR system. Under above optimal conditions, abundant, stable and clear strips could be obtained and the primer S1458 with good repeatability and polymorphism was further screened out. It was found that the specific fragment about 1500bp was observed in shuffled strain and partial sequence band had a high homology with the MarR-family transcription regulator factors existing in several streptomyces. The results indicated that RAPD technology was contribute to determining genotypic differences in Streptomyces gilvosporeus strains from genome shuffling and serving as an effective tool for exploring the biosynthetic genes correlated with metabolic product.