摘要
将前人报道的乳酸菌质粒提取方法与大肠杆菌质粒提取方法相结合,对常用试剂盒质粒提取方法进行改进,建立了一种快速、安全、高效的乳酸菌质粒提取方法。提取过程中,溶菌酶最佳浓度为20mg/ml,最佳处理时间为30min,同时避免了毒性物质溴化乙锭的使用。多次实验结果表明,采用改进后的方法可明显提高乳酸菌天然质粒的提取效果。且重复性好,为下一步乳酸菌的分子改良奠定了基础。
By combining the extraction method of plasmid from E.coil and the method of plasmid from lactic acid bacteria reported before to improve the common methods used in plasmid extraction kit,a fast,safe and high efficient method for extraction of plasmid DNA from lactic acid bacteria was developed.The optimal concentration of lysozyme was proved to be 20mg/ml,the optimal processing time was 30min,and the usage toxic ethidium bromide was avoided during the whole process of extraction.Experimental results showed that,the developed method showed an increased extraction efficiency,as well as good reproducibility,and thus provided a good foundation for the following molecular improvement of lactic acid bacteria.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2010年第2期94-99,共6页
China Biotechnology
基金
河南省高校杰出科研人才创新工程资助项目(2006KYCX008)
关键词
乳酸菌
天然质粒提取
方法优化
Iactic acid bacteria Extraction of plasmid Method optimization
