| 酿酒酵母SNF4基因敲除缺失菌株的构建 |
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| 引用本文： | 林晓华,柯崇榕,吴毕莎,郑永标,李力,陈由强,黄建忠. 酿酒酵母SNF4基因敲除缺失菌株的构建[J]. 生物工程学报, 2011, 27(4): 572-578 |
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| 作者姓名： | 林晓华 柯崇榕 吴毕莎 郑永标 李力 陈由强 黄建忠 |
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| 作者单位： | 1. 福建师范大学工业微生物教育部工程研究中心,生命科学学院,福建省现代发酵技术工程研究中心,福州,350108
2. 福建师范大学生命科学学院,农业部甘蔗遗传改良重点开放实验室,福州,350108 |
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| 基金项目： | 现代农业产业技术体系建设专项资金 (No. nycytx-024-01-20)，公益性行业 (农业) 科研专项项目 (No. nyhyzx07-019)，农业部948项目 (No. 2006-G37)，福建省发改委重大项目 (No. [2004]477)，福建省科技厅平台建设项目 (No. 2005Q007) 资助。 |
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| 摘 要： | 构建一株酿酒酵母SNF4基因缺失菌株并研究其对乙醇产量的影响。扩增带有SNF4基因上下游同源序列和Kanr筛选标记的SNF4基因敲除组件，转化到酿酒酵母YS2获得阳性克隆子，然后将质粒pSH65转到阳性克隆子中，半乳糖诱导pSH65表达Cre酶切除Kanr筛选标记，获得SNF4等位基因完全缺失菌株YS2-△SNF4。发酵实验结果表明，缺失菌株YS2-△SNF4乙醇产量较出发菌株提高了7.57％。利用Cre-LoxP系统，成功构建了SNF4等位基因完全缺失菌株并提高乙醇产生量。
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| 关 键 词： | 酿酒酵母，SNF4，基因敲除 |
| 收稿时间： | 2010-08-03 |
Construction of Saccharomyces cerevisiae mutant with knockout of SNF4 gene |
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| Xiaohua Lin,Chongrong Ke,Bisha Wu,Yongbiao Zheng,Li Li,Youqiang Chen and Jianzhong Huang. Construction of Saccharomyces cerevisiae mutant with knockout of SNF4 gene[J]. Chinese journal of biotechnology, 2011, 27(4): 572-578 |
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| Authors: | Xiaohua Lin Chongrong Ke Bisha Wu Yongbiao Zheng Li Li Youqiang Chen Jianzhong Huang |
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| Affiliation: | Engineering Research Center of Industrial Microbiology, Ministry of Education, Engineering Research Center of Fujian Modern Fermentation Technology, College of Life Sciences, Fujian Normal University, Fuzhou 350108, China;Engineering Research Center of Industrial Microbiology, Ministry of Education, Engineering Research Center of Fujian Modern Fermentation Technology, College of Life Sciences, Fujian Normal University, Fuzhou 350108, China;Engineering Research Center of Industrial Microbiology, Ministry of Education, Engineering Research Center of Fujian Modern Fermentation Technology, College of Life Sciences, Fujian Normal University, Fuzhou 350108, China;Engineering Research Center of Industrial Microbiology, Ministry of Education, Engineering Research Center of Fujian Modern Fermentation Technology, College of Life Sciences, Fujian Normal University, Fuzhou 350108, China;Engineering Research Center of Industrial Microbiology, Ministry of Education, Engineering Research Center of Fujian Modern Fermentation Technology, College of Life Sciences, Fujian Normal University, Fuzhou 350108, China;Key Laboratory of Sugarcane Genetic Improvement, Ministry of Agriculture, College of Life Sciences, Fujian Normal University, Fuzhou 350108, China;Engineering Research Center of Industrial Microbiology, Ministry of Education, Engineering Research Center of Fujian Modern Fermentation Technology, College of Life Sciences, Fujian Normal University, Fuzhou 350108, China |
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| Abstract: | Construction and ethanol production effects of SNF4 gene knockout in Saccharomyces cerevisiae were described in this paper.For knockout of SNF4 gene in S.cerevisiae YS2,a PCR-amplified disruption cassette was used,encoding the short flanking homologous regions to the SNF4 gene and Kanr as selectable marker.The SNF4 gene disruption cassette was transformed into S.cerevisiae YS2 through LiAc/SS Carrier DNA/PEG.The positive transformants were grown on G418 plates and verified by PCR.The Kanr marker was rescued... |
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| Keywords: | Saccharomyces cerevisiae SNF4 gene knockout |
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