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| 大肠杆菌植酸酶appA2基因真核表达载体的构建及在细胞中的表达 | |
| 引用本文： | 白立景,鞠辉明,牟玉莲,杨述林,李奎,陈明勇.大肠杆菌植酸酶appA2基因真核表达载体的构建及在细胞中的表达[J].中国兽医杂志,2011,47(7). |
| 作者姓名： | 白立景 鞠辉明 牟玉莲 杨述林 李奎 陈明勇 |
| 作者单位： | 1. 中国农业大学动物医学院,北京 海淀 100193;中国农业科学院北京畜牧兽医研究所,北京 海淀 100193 2. 中国农业科学院北京畜牧兽医研究所,北京 海淀,100193 3. 中国农业大学动物医学院,北京 海淀,100193 |
| 基金项目： | 国家自然科学基金项目(30830080); 国家转基因新品种培育重大专项(2008ZX08006-003) |
| 摘 要： | 根据已经公布的植酸酶appA2基因序列,设计合成了1对特异性引物,应用RT-PCR技术从大肠杆菌中扩增得到植酸酶appA2基因,并将其克隆到真核表达载体pcDNA3.1(+)中,构建了重组真核表达质粒pcDNA-appA2,对重组表达质粒鉴定正确后,转染猪PK15细胞,经G418筛选后,通过实时荧光定量PCR检测细胞内appA2表达,同时测定细胞内植酸酶的活性,检测结果表明,本试验成功构建了pcDNA-appA2重组真核表达载体,转染细胞后appA2的表达量是对照组的1 686.55倍,同时具有较好的植酸酶活性,为通过生物反应器制备植酸酶研究奠定了基础。 |
| 关 键 词： | 植酸酶 appA2基因 真核表达载体 表达 |
Construction of eukaryotic expression vector and expression of appA2 gene of Escherichia coli in PK15 |
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| BAI Li-jing,JU Hui-ming,MU Yu-lian,YANG Shu-lin,LI Kui,CHEN Ming-yong.Construction of eukaryotic expression vector and expression of appA2 gene of Escherichia coli in PK15[J].Chinese Journal of Veterinary Medicine,2011,47(7). | |
| Authors: | BAI Li-jing JU Hui-ming MU Yu-lian YANG Shu-lin LI Kui CHEN Ming-yong |
| Institution: | BAI Li-jing1,2,JU Hui-ming2,MU Yu-lian2,YANG Shu-lin2,LI Kui2,CHEN Ming-yong1(1.College of Veterinary Medicine,China Agricultural University,Beijing 100193,China,2.Institute of Animal Science,Chinese Academy of Agricultural Sciences,China) |
| Abstract: | According to the published sequence of Phytase appA2 gene,a pair of primers were designed and synthesized.Phytase appA2 gene was amplified by RT-PCR from Escherichia coli,and cloned into pcDNA3.1(+) vector.The eukaryotic expression vector pcDNA-appA2 was successfully constructed.Phytase appA2 gene was sequenced and compared with the published sequence of Phytase appA2 gene in GenBank.The expression of recombinant plasmid pcDNA-appA2 in PK15 cells was induced and detected by Quantitative fluorogenic real-tim... |
| Keywords: | phytase appA2 gene eukaryotic expression vector expression |
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