酒类酒球菌快速特异性PCR鉴定体系的优化  

Optimization of rapid specificity identification of Oenococcus oeni

作　　者：王华[1,2] 金刚[1] 李翠霞[3] 杜立业[1] 李华[1,2] 

机构地区：[1]西北农林科技大学葡萄酒学院,陕西杨凌712100 [2]陕西省葡萄与葡萄酒工程技术研究中心,陕西杨凌712100 [3]西北农林科技大学生命科学学院,陕西杨凌712100

出　　处：《中国酿造》2010年第5期152-156,共5页China Brewing

基　　金：农业部"948"项目(2009-Z29)

摘　　要：研究采用目前应用较多的快速鉴定方法-种属特异性PCR技术(species-specific PCR)对酒类酒球菌进行鉴定。比较了不同提取方法所提DNA模板对species-specific PCR扩增结果的影响。尝试通过优化反应体系和反应条件,直接使用菌液和菌落作为模板进行扩增。结果表明,使用优化后的方法提取的DNA不需要纯化就能得到稳定的扩增结果,在优化后的反应体系下,可直接使用菌液或菌落作为模板进行扩增,扩增结果稳定,条带清晰。The research on classification and identification of Oenococcus oeni in China is still fragmentary. In order to establish a fast, reliable and e- conomical identification system, we referred to a pair of specific primers of O. oerd to do species-specific PCR. For the purpose of finding suitable re- action conditions, we compared the PCR amplification results of different DNA templates that extracted by CTAB and lysozyme methods, And we tried to use the bacterial suspension and colonies as DNA template for direct PCR amplification. The results showed that the DNA template extracted by our method needn＇t to be purified, and it can be used to PCR amplification directly. With the optimized reaction system, bacterial suspension or colonies can be used directly as DNA template to amplify the malolactic enzyme gene of O. oeni by PCR, the result is stable. The method is simple, fast, and reliable, ane it is suitable for rapid identification of O. oeni.

关 键 词：酒类酒球菌 种属特异性PCR鉴定 苹果酸-乳酸酶基因 快速鉴定 

分 类 号：TS261.1[轻工技术与工程—发酵工程]