乳酸乳球菌质粒提取方法的改进与优化  

Improvement and Optimization of A Method for Extraction of Plasmid from Lactococcus lactis

作　　者：任志娟[1] 李轶杰[1] 马正海[1] 

机构地区：[1]新疆生物资源基因工程重点实验室,新疆大学生命科学与技术学院,乌鲁木齐830046

出　　处：《新疆农业科学》2011年第4期734-738,共5页Xinjiang Agricultural Sciences

基　　金：国家自然科学基金项目(30860265);新疆生物资源基因工程重点实验室基金(XJDX0201-2010-2)

摘　　要：【目的】建立一种简便、快速、安全的乳酸乳球菌质粒提取方法。【方法】质粒提取采用文献报道的乳酸菌质粒提取与常用的大肠杆菌质粒提取相结合的方法,并对提取过程中溶菌酶浓度、溶菌酶处理方式和时间进行优化。【结果】采用改良后的方法提取乳酸菌质粒效果显著。最佳溶菌酶浓度是10 mg/mL,最佳溶菌酶处理方式和时间是37℃振荡培养30 min。【结论】该方法避免了毒性物质溴化乙锭的使用,同时减少了通常提取乳酸菌质粒的复杂步骤,是一种高效、快速、安全的提取乳酸菌质粒的方法。【Objective】 To develop a simple,rapid and safe method for extraction of plasmid DNA from Lactococcus lactis.【Method】The method for plasmid extraction from lactic acid bacteria was improved by combining the method of E.coli plasmid extraction and the reported method of plasmid extraction from lactic acid bacteria（LAB）.Conditions for this new method were optimized in terms of lysozyme concentration and lysozyme treatment way and time length..【Result】 The optimal concentration of lysozyme was 10 mg/ml,the optimal lysozyme treatment method and time were at 37℃ for 30 min with shaking.【Conclusion】This method avoided the use of ethidium bromide（EB） and simplified the complicated procedure,so it was a high effective and safe LAB plasmid DNA extraction method.

关 键 词：乳酸菌 质粒 提取 溶菌酶 

分 类 号：S188.4[农业科学—农业基础科学]