| 家蚕磷酸吡哆醇氧化酶的体外定点突变及其活性鉴定 |
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| 引用本文： | 童宁,张剑韵,黄龙全. 家蚕磷酸吡哆醇氧化酶的体外定点突变及其活性鉴定[J]. 昆虫学报, 2011, 54(11): 1231-1235 |
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| 作者姓名： | 童宁 张剑韵 黄龙全 |
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| 作者单位： | 1. 安徽农业大学茶与食品科技学院,合肥,230036
2. 安徽农业大学生命科学院,合肥,230036 |
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| 基金项目： | 国家自然科学基金项目(30870338) |
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| 摘 要： | [目的]研究家蚕Bombyx mori磷酸吡哆醇氧化酶(pyridoxine 5’-phosphate oxidase,PNPO)个别保守氨基酸残基对PNPO酶活性的影响.[方法]用重叠延伸法把氨基酸残基Lys111 (AAA)突变为Glu (GAA),Ser160(AGC)定点突变为Ala(GCC)；构建重组表达载体...
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| 关 键 词： | 家蚕 磷酸吡哆醇氧化酶 定点突变 基因表达 蛋白纯化 酶活性 |
Site-directed mutation of pyridoxine 5'-phosphate oxidase from Bombyx mori and activity assay of the mutants in vitro |
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| TONG Ning , ZHANG Jian-Yun , HUANG Long-Quan. Site-directed mutation of pyridoxine 5'-phosphate oxidase from Bombyx mori and activity assay of the mutants in vitro[J]. Acta Entomologica Sinica, 2011, 54(11): 1231-1235 |
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| Authors: | TONG Ning ZHANG Jian-Yun HUANG Long-Quan |
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| Affiliation: | TONG Ning~1,ZHANG Jian-Yun~2,HUANG Long-Quan~1,*(1.College of Tea & Food Science,Anhui Agricultural University,Hefei 230036,China,2.College of Life Science,China) |
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| Abstract: | [Aim] To study the effects of single conserved amino acid residue of pyridoxine 5'-phosphate oxidase(PNPO) from Bombyx mori on its activity.[Methods] Lys111(AAA) and Ser160(AGC),two of the most conserved residues,were mutated to Glu(GAA) and Ala(GCC) by using over-lap extension,respectively.The obtained expression plasmids were over-expressed in Escherichia coli Rosetta by IPTG induction,and then the expressed products were purified with affinity chromatography and the activity of the purified PNPO were det... |
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| Keywords: | Bombyx mori pyridoxine 5'-phosphate oxidase site-directed mutation gene expression protein purification enzyme activity |
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