| 黑曲霉A9葡萄糖氧化酶基因克隆及其酵母表达载体构建 |
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| 引用本文： | 黄亮,郭润芳,于宏伟,贾英民.黑曲霉A9葡萄糖氧化酶基因克隆及其酵母表达载体构建[J].河北农业大学学报,2008,31(1):60-64. |
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| 作者姓名： | 黄亮 郭润芳 于宏伟 贾英民 |
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| 作者单位： | 河北农业大学,食品科技学院,河北,保定,071001 |
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| 基金项目： | 河北省“十一五”科技攻关项目资助(农产品精深加工用主要微生物菌种培育及菌种资源库建设06220106D) |
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| 摘 要： | 以黑曲霉A9基因组为模板,利用聚合酶链式反应(PCR)直接对葡萄糖氧化酶(GOD)基因进行PCR扩增,再将PCR产物克隆入pMD18-T载体,经DNA测序,该基因为1 743 bp,编码581个氨基酸。该基因序列已在GeneBank注册,登录号为DQ836361。用限制性内切酶切下目的基因,插入到毕赤酵母表达载体pPIC9K中,构建成表达载体pPIC9K-GOD。重组毕赤酵母表达载体pPIC9K-GOD的构建,可望获得超量表达的高活性葡萄糖氧化酶,为研制高品质的酶制剂奠定基础。
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| 关 键 词： | 黑曲霉 葡萄糖氧化酶 克隆 表达载体pPIC9K |
| 文章编号： | 1000-1573(2008)01-0060-05 |
| 修稿时间： | 2007年3月9日 |
Cloning of Aspergillus niger A9 glucose oxidase gene and construction of the expression vector |
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| HUANG Liang,GUO Run-fang,YU Hong-wei,JIA Ying-min.Cloning of Aspergillus niger A9 glucose oxidase gene and construction of the expression vector[J].Journal of Agricultural University of Hebei,2008,31(1):60-64. |
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| Authors: | HUANG Liang GUO Run-fang YU Hong-wei JIA Ying-min |
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| Abstract: | The DNA fragment encoding Aspergillus niger A9 glucose oxidase was amplified by PCR using A.niger A9 genomic DNA as template.Then the product of PCR was inserted into vector pMD18-T.The sequence analysis showed that the cloned DNA had a length of 1 743 bp,encoding a protein of 581 amino acids.(GenBank Accession number: DQ836361).After digested by both NotⅠand AvrⅡ,DNA fragment was linked with vector plasmid pPIC9K by T4 DNA ligase.The construction of expression vector pPIC9K-GOD may lead to obtaining large glucose oxidase,thus providing basis for further research on developing high quality enzymes. |
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| Keywords: | Aspergillus niger glucose oxidase clone expression vector pPIC9K |
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