基于定量PCR方法初步分析西藏开菲尔粒中细菌与酵母菌数量变化

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基于定量PCR方法初步分析西藏开菲尔粒中细菌与酵母菌数量变化
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作者:
                        王兴兴王兴兴
上海海洋大学 食品学院, 上海 201306
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丁梦帆丁梦帆
上海海洋大学 食品学院, 上海 201306
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潘迎捷潘迎捷
上海海洋大学 食品学院, 上海 201306;上海水产品加工及贮藏工程技术研究中心, 上海 201306;农业部水产品贮藏保鲜质量安全风险评估实验室, 上海 201306
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王永杰王永杰
上海海洋大学 食品学院, 上海 201306;上海水产品加工及贮藏工程技术研究中心, 上海 201306;农业部水产品贮藏保鲜质量安全风险评估实验室, 上海 201306
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作者单位:上海海洋大学,上海海洋大学,上海海洋大学食品学院,上海海洋大学食品学院
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基金项目:上海高校特聘教授东方学者岗位计划基金(20101222);上海市科学技术委员会科技支撑项目(10540503000);上海市科委工程中心建设(11DZ2280300)

The number of bacteria and yeasts in Tibetan kefir grains monitored by real-time qPCR

Author:

WANG Xingxing
WANG Xingxing
College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China
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DING Mengfan
DING Mengfan
College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China
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PAN Yingjie
PAN Yingjie
College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China;Shanghai Engineering Research Center of Aquatic-Product Processing & Preservation, Shanghai 201306, China;Laboratory of Quality & Safety Risk Assessment for Aquatic Product on Storage and Preservation, Ministry of Agriculture, Shanghai 201306, China
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WANG Yongjie
WANG Yongjie
College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China;Shanghai Engineering Research Center of Aquatic-Product Processing & Preservation, Shanghai 201306, China;Laboratory of Quality & Safety Risk Assessment for Aquatic Product on Storage and Preservation, Ministry of Agriculture, Shanghai 201306, China
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Affiliation:

shanghai ocean university,shang hai ocean university,shanghai ocean university,shang hai ocean university

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摘要:

运用实时荧光定量PCR方法分析西藏开菲尔粒中细菌与酵母菌的数量变化规律, 需要有效提取其微生物总基因组DNA。本文通过同时添加溶菌酶与溶壁酶优化实验方案, 成功获得西藏开菲尔粒中微生物的总基因组DNA。荧光定量PCR分析提取自培养1、4、8和12周的西藏开菲尔粒的微生物总基因组DNA, 结果显示:4个时间点细菌16S rRNA与酵母菌26S rRNA基因拷贝数均分别约为每微升10⁷拷贝与10⁵拷贝, 表明细菌与酵母菌的数量相对稳定。该结果为进一步研究西藏开菲尔粒中细菌与酵母菌间的共生关系奠定了一定的基础。

关键词:西藏开菲尔粒;基因组提取;实时荧光定量PCR;数量变化

Abstract:

In order to understand whether the total number of bacteria and yeasts in Tibetan kefir grains was subjected to change by using qPCR method, their genomic DNAs were prepared successfully by adding lysozyme and lyticase simultaneously. Subsequently, qPCR was performed. It revealed that the gene copy number of bacteria (16S rRNA gene) and yeasts (26S rRNA gene) was 10⁷ copies/μL and 10⁵ copies/μL respectively after 1, 4, 8 and 12 week(s) of cultivation, which indicates that the communities of bacteria and yeasts are relatively stable in Tibetan kefir grains. These results give an insight into exploration of symbiosis mechanism between bacteria and yeasts in Tibetan kefir grains.

Key words:Tibetan kefir grains;DNA extraction;real-time qPCR;changing of cell numbers

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引用本文

王兴兴,丁梦帆,潘迎捷,王永杰.基于定量PCR方法初步分析西藏开菲尔粒中细菌与酵母菌数量变化[J].上海海洋大学学报,2015,24(4):625-631.
WANG Xingxing, DING Mengfan, PAN Yingjie, WANG Yongjie. The number of bacteria and yeasts in Tibetan kefir grains monitored by real-time qPCR[J]. Journal of Shanghai Ocean University,2015,24(4):625-631.

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收稿日期:2014-12-23
最后修改日期:2015-04-17
录用日期:2015-05-22
在线发布日期: 2015-07-22
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