| 牛对氧磷酶1基因和对氧磷酶2基因原核表达载体的构建及其表达分析 |
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| 引用本文： | 姬爱国,淮亚红,张路培,李俊雅,王淑辉,许尚忠,高雪,任红艳.牛对氧磷酶1基因和对氧磷酶2基因原核表达载体的构建及其表达分析[J].中国畜牧兽医,2008,35(9):59-63. |
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| 作者姓名： | 姬爱国 淮亚红 张路培 李俊雅 王淑辉 许尚忠 高雪 任红艳 |
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| 作者单位： | 1. 中国农业科学院北京畜牧兽医研究所，北京 100193; 2.西北农林科技大学动物科技学院，杨凌 712100 |
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| 基金项目： | 国家高技术研究发展计划(863计划)
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国家科学技术支撑计划
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| 摘 要： | 为制备和纯化对氧磷酶(PON1和PON2)蛋白做准备，对牛的PON1和PON2基因进行克隆并在大肠杆菌中表达。从牛肝脏组织中用RT-PCR扩增PON1和PON2 cDNA的全长编码序列，连接到原核表达载体pET28a中，构建重组表达质粒并转化到大肠杆菌BL21（DE3）中，以IPTG诱导表达目的蛋白，SDS-PAGE分析表达情况。 结果表明，成功克隆了PON1和PON2两个基因的cDNA的全长编码序列，酶切及测序鉴定证明，获得含有目的基因片段的重组质粒，表达的牛的PON1和PON2融合蛋白以可溶性的形式存在。这为进一步制备和纯化PON1和PON2蛋白，以及探索其在家畜使用寿命中的研究和提高犊牛初生重的作用等方面提供依据。
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| 关 键 词： | 对氧磷酶 对氧磷酶1基因 对氧磷酶2基因 克隆 原核表达 |
Bovine PON1 Gene and PoN2 Gene Cloning and Prokaryotic Expression |
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| JI Ai-guo,HUAI Ya-hong,ZHANG Lu-pei,LI Jun-ya,WANG Shu-hui,XU Shang-zhong,GAo Xue,REN Hong-yan.Bovine PON1 Gene and PoN2 Gene Cloning and Prokaryotic Expression[J].China Animal Husbandry & Veterinary Medicine,2008,35(9):59-63. |
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| Authors: | JI Ai-guo HUAI Ya-hong ZHANG Lu-pei LI Jun-ya WANG Shu-hui XU Shang-zhong GAo Xue REN Hong-yan |
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| Institution: | 1. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China;2. College of Animal Science and Technology, Northwest Sci Tech University of Agriculture and Forestry, Yangling 712100, China |
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| Abstract: | To construct and express prokaryotic expression vector of bovine paraoxonase-1 and paraoxonase-2 (PON1 and PON2) and to obtained pure PON1 and PON2. Full length sequences of PON1 and PON2 genes were amplified from bovine liver by RT-PCR and were cloned into the expression vector--pET28a. PON1 and PON2 fusion proteins were expressed in BL21 under IPTG induction and the expressed fusion proteins were detected by SDS-PAGE. The recombinant plasmids containing the target genes were constructed successfully. The fusion proteins were expressed in E coli in soluble form. The fusion proteins are expressed in soluble form, which is a useful reagent for further preparation of purifying and study of the function of PON1 and PON2 in blood vessel and diabetes mellitus, and boosting the birth weight of calves. |
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| Keywords: | paraoxonases PON1 gene PON2 gene clone prokaryotic expression |
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