| 酿酒酵母HOR2基因缺失突变株的构建 |
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| 引用本文： | 陈永金,林晓华,王艳尊,雷娟娟,黄建忠. 酿酒酵母HOR2基因缺失突变株的构建[J]. 生物技术, 2010, 20(2): 6-8 |
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| 作者姓名： | 陈永金 林晓华 王艳尊 雷娟娟 黄建忠 |
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| 作者单位： | 福建师范大学生命科学学院,工业微生物教育部工程研究中心,福建现代发酵技术工程研究中心,福建福州350108 |
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| 基金项目： | 国家"948"项目，福建省发改委重大项目，福建省科技平台建设项目 |
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| 摘 要： | 目的:构建酿酒酵母HOR2基因缺失的突变株并研究其对甘油和乙醇产量的影响。方法:以PCR为基础,通过同源重组的方式使目的基因缺失。结果:通过设计含有与HOR2(GPP2)基因两侧序列同源的长引物,以质粒PUG6为模板进行PCR构建含有Cre/loxP系统的酿酒酵母HOR2基因敲除组件,转化酿酒酵母(Saccharomyces cerevisiae)YS2,获得为loxP-kan-loxP序列组件所替换而产生kanr的阳性克隆子。然后再将质粒PSH65转入阳性克隆子诱导表达Cre酶切除筛选标记,在原ORF基因处保留一个loxP位点,丢失质粒后获得HOR2单倍体缺陷型菌株。重复转化敲除组件实现另一条等位基因的敲除。发酵实验表明,突变株甘油产量降低3.34%,乙醇产量提高1.96%。结论:成功获得了酿酒酵母HOR2基因缺失的突变株,并命名为YS2-HOR2。
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| 关 键 词： | 酿酒酵母 HOR2基因 基因敲除 乙醇 甘油 |
Construction of S. cerevisiae Mutants Deleted HOR2 Gene |
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| CHEN Yong-jin,LIN Xiao-hua,WANG Yan-zun,LEI Juan-juan,HUANG Jian-zhong. Construction of S. cerevisiae Mutants Deleted HOR2 Gene[J]. Biotechnology, 2010, 20(2): 6-8 |
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| Authors: | CHEN Yong-jin LIN Xiao-hua WANG Yan-zun LEI Juan-juan HUANG Jian-zhong |
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| Affiliation: | CHEN Yong-jin,LIN Xiao-hua,WANG Yan-zun,LEI Juan-juan,HUANG Jian-zhong(Engineering Research Center of Industrial Microbiology,Ministry of Education,College of Life Science,Fujian Normal University,Fuzhou 350108,China) |
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| Abstract: | Objective:Construction of S.cerevisiae mutants deleted HOR2 gene and studies on its effect on biosynthesis of ethanol and glycerol in Saccharomyces cerevisiae.Method:Disrupting HOR2 gene by means of homologous recombination.Result:The gene disruption cassette produced by PCR using the same long oligonucleotides which comprise 19 or 22 3' nucleotides complementary to sequences in the templates(pUG6 marker plasmid)and 45 5' nucleotides that annealed to sites upstream or downstream of the genomic target sequen... |
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| Keywords: | Saccharomyces cerevisiae HOR2 gene gene knockout ethanol glycerol |
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