高效检测分析玫瑰黄链霉菌Men-myco-93-63线性质粒

Efficient detection and analysis of linear plasmid in Stremptomyces Men-myco-93-63

摘要:

【目的】建立一种高效检测和分析玫瑰黄链霉菌Men-myco-93-63内源质粒的方法。【方法】根据“链霉菌线性质粒5′末端都结合有共价蛋白”这一性质，通过改进琼脂糖包埋块制作方法，发明了一项快速高效检测链霉菌质粒种类、构型及大小的技术，称为“包埋块二次处理法”，该方法还能从正反两方面表明共价蛋白的存在与否。【结果】利用此方法高效检测出了玫瑰黄链霉菌Men-myco-93-63中存在两种线性质粒，双向电泳验证了该方法的正确性，推测质粒大小分别约为 47 kb和53 kb。【结论】利用本研究发明的方法首次成功地检测了玫瑰黄链霉菌Men-myco-93-63中的内源线性质粒。

Abstract:

[Objective] To develop an efficient method to detect and analyze the endogenous plasmid in Stremptomyces Men-myco-93-63. [Methods] According to the property of ‘the linear chromosomes and plasmids of Stremptomyces spp.have proteins covalently bound to the 5′ends of the DNA′, a novel method was established to detect the type, configuration and size of the Streptomyces spp. plasmids by improving the preparation of agarose embedded block. The method can also show whether the linear plasmids have proteins covalently bound to the DNA from both the positive and negative aspects. [Results]The type and configuration of two plasmids were determined efficiently from Stremptomyces roseoflavus Men-myco-93-63 and verified by the two-dimensional electrophoresis. The speculated sizes of these plasmids were 47 kb and 53 kb, respectively. [Conclusion] Two linear plasmids were detected successfully from Stremptomyces roseoflavus Men-myco-93-63 by a new method.