使用MseI限制性内切酶对放线菌链霉菌属中的12株菌基因组DNA进行酶切,与接头连接后,引物使用一个选择性碱基对模板进行PCR扩增,PCR产物在琼脂糖凝胶上进行电泳检测。结果表明,对于MseI内切酶产生的模板,选择性碱基采用A、T、C和G都能够获得清晰丰富的条带。MseI内切酶产生的图谱上有72个位点,多态性位点71个,达98.6%。因此,使用MseI内切酶适合构建放线菌的单酶切AFLP指纹图谱。
Whole genomic DNA of 12 Streptomyces strains were digested with restriction enzyme MseI. After being ligated with MseI adaptor, restriction fragments were amplified with primers containing only one arbitrarily chosen base. AFLP-PCR pro ducts were separated with agarose gel electrophoresis. The result showed that clear and rich polymorphism bands were obtained when the chosen base was A, T, C or G. Cluster analysis showed that each species located in different branch of the phylogeny. The method is easy, low-cost, high-resolution and high-throughout.