| 猪尿酸氧化酶基因的克隆及表达 |
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| 引用本文： | 薛璟,汪维云,陈建华.猪尿酸氧化酶基因的克隆及表达[J].安徽农业大学学报,2011,38(4):592-595. |
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| 作者姓名： | 薛璟 汪维云 陈建华 |
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| 作者单位： | 安徽农业大学生命科学学院,合肥,230036;中国药科大学生命科学与技术学院,南京,210009 |
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| 摘 要： | 本研究主要涉及猪尿酸氧化酶(urate oxidase,EC 1.7.3.3)的原核表达载体的构建及蛋白表达,并对其酶活性进行测定。从猪肝组织细胞中提取总的RNA,利用RT-PCR技术克隆pUOX(porcine urate oxidase),插入原核表达载体pET-22b中,构建重组表达载体pET-22b/pUOX,并转化到大肠杆菌(Escherichia coli)BL21(DE3)中。通过IPTG诱导获得的重组蛋白经SDS-PAGE检测,在约34 ku处有一清晰蛋白条带,这与预期分子量一致。测定该重组蛋白的酶活力达到1.962 U,这为后期实验奠定了重要的基础。
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| 关 键 词： | 猪尿酸氧化酶 基因克隆 表达 |
Cloning and expression of porcine urate oxidase |
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| XUE Jing,WANG Wei-yun and CHEN Jian-hua.Cloning and expression of porcine urate oxidase[J].Journal of Anhui Agricultural University,2011,38(4):592-595. |
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| Authors: | XUE Jing WANG Wei-yun and CHEN Jian-hua |
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| Institution: | 1.School of Life Science,Anhui Argicultural University,Hefei 230036; 2.School of Life Science and Biotechnology,China Pharmaceutical University,Nanjing 210009) |
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| Abstract: | This study is aimed at constructing of prokaryotic expression vector to express of porcine urate oxidase (urate oxidase, EC 1.7.3.3), and determine its activity. Total RNA was extracted from pig liver, and pUOX (porcine urate?oxidase) gene was cloned by RT-PCR technology; the gene was inserted into the prokaryotic expression vector pET-22b to construct the recombinant expression vector pET-22b/pUOX, and then transformed into Escherichia coli?BL21 (DE3).?Recombinant proteins, collected with IPTG induction, were detected to show a clear protein band about 34 ku by SDS-PAGE, which is consistent with the expected molecular weight. The enzyme activity of recombinant protein was 0.392 U, which?laid an important foundation for farther research. |
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| Keywords: | porcine urate oxidase cloning expression |
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