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6种区分乳酸乳球菌乳酸亚种和乳酸乳球菌乳脂亚种的分子生物学方法比较
引用本文:张家超,王芳,徐海燕,于洁,刘文俊,包秋华,孙志宏,张和平.6种区分乳酸乳球菌乳酸亚种和乳酸乳球菌乳脂亚种的分子生物学方法比较[J].微生物学报,2010,50(12):1670-1676.
作者姓名:张家超  王芳  徐海燕  于洁  刘文俊  包秋华  孙志宏  张和平
作者单位:内蒙古农业大学乳品生物技术与工程教育部重点实验室,呼和浩特,010018
基金项目:国家自然基金项目 (30660135,30800861);国家863项目(2006AA10Z345,2007AA10Z353)
摘    要:【目的】比较并评价6种分子生物学技术对乳酸乳球菌乳酸亚种(Lactococcus lactis subsp.lactis)和乳酸乳球菌乳脂亚种(Lactococcus lactis subsp.cremoris)的区分效果。【方法】采用16S rRNA基因序列分析技术,16S-23S rRNA间区序列多态性分析技术,变性梯度凝胶电泳技术(DGGE),随机扩增多态性分析技术(RAPD),重复基因外回文序列分析技术(rep-PCR)和限制性酶切片段多态性分析技术(RFLP)对4株Lactococcus lactis subsp.lactis和Lactococcus lactis subsp.cremoris参考菌株进行了区分,并对这6种方法的区分效果进行了比较评价。【结果】16S rRNA基因序列分析技术,16S-23S rRNA间区序列多态性分析技术无法区分Lactococcus lactis subsp.lactis和Lactococcus lactis subsp.cremoris,而其余4种技术可以实现区分。【结论】变性梯度凝胶电泳(DGGE),随机扩增多态性分析技术(RAPD)耗时短,操作简单,试验结果准确稳定,更适合Lactococcus lactis subsp.lactis和Lactococcus lactis subsp.cremoris的快速准确区分。

关 键 词:关键词:乳酸乳球菌  分类鉴定  分子生物学方法
收稿时间:6/3/2010 12:00:00 AM

Comparison of Six Molecular Methods for Differentiation of Lactococcus lactis subsp. lactis and cremoris
Jiachao Zhang,Fang Wang,Haiyan Xu,Jie Yu,Qiuhua Bao,Wenjun Liu,Zhihong Sun and Heping Zhang.Comparison of Six Molecular Methods for Differentiation of Lactococcus lactis subsp. lactis and cremoris[J].Acta Microbiologica Sinica,2010,50(12):1670-1676.
Authors:Jiachao Zhang  Fang Wang  Haiyan Xu  Jie Yu  Qiuhua Bao  Wenjun Liu  Zhihong Sun and Heping Zhang
Institution:Key Laboratory of Dairy Biotechnology and Bioengineering, Ministry of Education , Department of Food Science and Engineering, Inner Mongolia Agricultural University, Hohhot 010018, China;Key Laboratory of Dairy Biotechnology and Bioengineering, Ministry of Education , Department of Food Science and Engineering, Inner Mongolia Agricultural University, Hohhot 010018, China;Key Laboratory of Dairy Biotechnology and Bioengineering, Ministry of Education , Department of Food Science and Engineering, Inner Mongolia Agricultural University, Hohhot 010018, China;Key Laboratory of Dairy Biotechnology and Bioengineering, Ministry of Education , Department of Food Science and Engineering, Inner Mongolia Agricultural University, Hohhot 010018, China;Key Laboratory of Dairy Biotechnology and Bioengineering, Ministry of Education , Department of Food Science and Engineering, Inner Mongolia Agricultural University, Hohhot 010018, China;Key Laboratory of Dairy Biotechnology and Bioengineering, Ministry of Education , Department of Food Science and Engineering, Inner Mongolia Agricultural University, Hohhot 010018, China;Key Laboratory of Dairy Biotechnology and Bioengineering, Ministry of Education , Department of Food Science and Engineering, Inner Mongolia Agricultural University, Hohhot 010018, China;Key Laboratory of Dairy Biotechnology and Bioengineering, Ministry of Education , Department of Food Science and Engineering, Inner Mongolia Agricultural University, Hohhot 010018, China
Abstract:Abstract: Objective] To compare six molecular methods for differentiation of Lactococcus lactis subsp. lactis and cremoris. Methods] Six molecular methods, such as 16S rRNA gene analysis, 16S-23S rRNA Intergenic spacer region polymorphism, Denaturing Gradient Gel Electrophoresis (DGGE), Random Amplified Polymorphic DNA (RAPD), Repetitive Extragenic Palindromic-PCR and Restricted Fragment Length Polymorphisms were used to differentiate the reference strains. Results] Except for 16S rRNA gene analysis and 16S-23S rRNA Intergenic spacer region polymorphism, the other protocols were competent. Conclusion] The methods of DGGE and RAPD were more appropriate for differentiation of Lactococcus lactis subsp. lactis and cremoris.
Keywords:Key words: Lactococcus lactis  differentiation  molecular methods
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