利用S1核酸酶对链霉菌进行DNA同源性研究。

DNA homology studies in Streptomyces using S1 nuclease.

作者信息

Furumai T, Sudoh M, Sawairi S, Maruyama H B

出版信息

J Antibiot (Tokyo). 1984 Jun;37(6):641-5. doi: 10.7164/antibiotics.37.641.

DOI:10.7164/antibiotics.37.641

Abstract

The optimal reaction conditions for the determination of DNA-homology in Streptomyces species were established in the presence of formamide using S1 nuclease. The melting temperature of Streptomyces DNA was 90 degrees C in 0.42 M NaCl containing 20% formamide in which the denaturation was completed by boiling for 5 minutes. In the S1 reaction mixture consisting of 5 U of S1 nuclease, 0.168 M NaCl, 1 mM ZnSO4 and 8% formamide at pH 4.8, single-stranded DNA was hydrolyzed by more than 98%, while the hydrolysis of double-stranded DNA was less than 3%. From the analysis of homoduplex formation, the C0t1/2 was found at 20 hours, when a mixture of unlabeled DNA and index DNA was used at a ratio of 500:1.

摘要

利用S1核酸酶在甲酰胺存在的情况下，建立了测定链霉菌属物种中DNA同源性的最佳反应条件。在含有20%甲酰胺的0.42M NaCl中，链霉菌DNA的解链温度为90℃，通过煮沸5分钟可完成变性。在由5U S1核酸酶、0.168M NaCl、1mM ZnSO4和pH 4.8的8%甲酰胺组成的S1反应混合物中，单链DNA的水解率超过98%，而双链DNA的水解率小于3%。通过对同源双链体形成的分析，当未标记DNA和指数DNA的混合物以500:1的比例使用时，在20小时时发现C0t1/2。