| 益生菌FGM葡萄糖激酶基因（glcK）在黄芪发酵中的表达 |
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| 引用本文： | 郝桂娟,王 龙,张 凯,张景艳,王学智,孟嘉仁,杨志强,李建喜.益生菌FGM葡萄糖激酶基因（glcK）在黄芪发酵中的表达[J].西北农业学报,2013,22(3):1-5. |
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| 作者姓名： | 郝桂娟 王 龙 张 凯 张景艳 王学智 孟嘉仁 杨志强 李建喜 |
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| 作者单位： | （中国农业科学院 兰州畜牧与兽药研究所，甘肃省中兽药工程技术研究中心， 甘肃省新兽药工程重点实验室，农业部兽用药物创制重点实验室，兰州 730050） |
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| 基金项目： | 国家自然科学基金（31072162）；国家行业农业科研专项(201303040)；公益性研究所基本科研费(1610322011007)。 |
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| 摘 要： | 旨在分析益生菌FGM在黄芪发酵过程中的生长动态，为阐述其作用机理奠定基础。采用平板计数法对益生菌FGM在黄芪发酵中的生长动态进行检测，同源克隆法克隆糖酵解途径中的限速酶葡萄糖激酶glcK基因序列，并运用SYBR Green Ⅰ实时荧光定量PCR技术分析其在黄芪发酵不同阶段的表达水平。结果表明：成功克隆得到glcK基因一段382 bp的片段（GenBank登录号JX976291），其序列一致性为83%～85%。实时荧光定量PCR结果显示，在FGM发酵6 h内，glcK基因表达上调且呈逐渐上升趋势，至对数期后期6 h表达水平达到最高（4.63倍）；稳定期初期8 h时表达水平显著下降（P<0.05），10 h至发酵结束基因表达下调。可见：在黄芪发酵6 h内FGM处于对数生长期，葡萄糖经糖酵解途径代谢产生ATP供细菌生长代谢利用，10 h后开始进入稳定期并进行次级代谢。
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| 关 键 词： | 黄芪 非解乳糖链球菌 发酵 葡萄糖激酶 基因克隆 实时荧光定量PCR |
Expression of glcK Gene Derived from the Probiotic FGM in Astragalus membranaceus Fermentation |
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| HAO Guijuan,WANG Long,ZHANG Kai,ZHANG Jingyan,WANG Xuezhi,MENG Jiaren,YANG Zhiqiang and LI Jianxi.Expression of glcK Gene Derived from the Probiotic FGM in Astragalus membranaceus Fermentation[J].Acta Agriculturae Boreali-occidentalis Sinica,2013,22(3):1-5. |
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| Authors: | HAO Guijuan WANG Long ZHANG Kai ZHANG Jingyan WANG Xuezhi MENG Jiaren YANG Zhiqiang and LI Jianxi |
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| Abstract: | This study was aimed at analyzing the growth of probiotic FGM in Astragalus membranaceus fermentation to lay a foundation for understanding the mechanism. FGM growth dynamics in A.membranaceus fermentation were investigated with plate count method. The nucleotide sequence of glucokinase catalyzing the rate limiting step of the glycolysis pathway was cloned and the variations of glcK gene expression during fermentation process was further analyzed by SYBR Green real time PCR. As a result, a segment of 382 bp (GenBank accession No. JX976291), displaying high sequence identity from 83% to 85%, was successfully obtained for the first time. Real time PCR results showed that glcK gene expression was up regulated and increased gradually within 6 h during A. fermentation. The expression level was peaked at 6 h and declined significantly at 8 h, and thereafter glcK gene expression was down regulated from 10 h to the end. Those results demonstrated that FGM was in exponential growth phase within 6 h, when glucose was metabolized for bacteria growth by the glycolysis pathway metabolism. After 10 h of fermentation, FGM was in stable phase for secondary metabolism. |
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| Keywords: | Astragalus membranaceus Streptococcus alactolyticus Fermentation Glucokinase Gene cloning Real time PCR |
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