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| 奶牛LAP基因真核表达载体的构建及其在COS-7细胞中的瞬时表达 | |
| 引用本文： | 曹随忠,袁曦,栾红雨,余树民,刘宗平.奶牛LAP基因真核表达载体的构建及其在COS-7细胞中的瞬时表达[J].中国兽医学报,2011,31(4). |
| 作者姓名： | 曹随忠 袁曦 栾红雨 余树民 刘宗平 |
| 作者单位： | 1. 四川农业大学动物医学院,四川,雅安,625014;扬州大学兽医学院,江苏扬州225009 2. 四川农业大学动物医学院,四川,雅安,625014 3. 扬州大学兽医学院,江苏扬州,225009 |
| 基金项目： | 中国博士后科学基金资助项目(20090451250); 扬州大学博士后基金资助项目(2008); 四川省教育厅青年基金资助项目(09ZB054) |
| 摘 要： | 根据GenBank上发表的牛舌抗菌肽(Lingual antimicrobial peptide,LAP)基因cDNA序列设计1对特异引物,从患乳腺炎的奶牛乳腺组织中以RT-PCR方法扩增LAP基因片段,将其克隆到pMD19-T Simple载体中测序。重组质粒经EcoRⅠ+BamHⅠ双酶切回收目的基因片段,亚克隆入增强型绿色荧光蛋白表达载体pEGFP-C1中构建重组融合表达质粒pEGFP-LAP,将其脂质体转染COS-7细胞,经荧光显微镜观察到融合表达的绿色荧光蛋白,采用RT-PCR检测到LAP基因在COS-7细胞中转录。pEGFP-LAP重组表达质粒的成功构建为进一步研究奶牛LAP基因的表达特性及利用基因工程技术防治奶牛乳腺炎奠定了基础。 |
| 关 键 词： | 奶牛 β-防御素 LAP基因 绿色荧光蛋白 真核表达 |
Construction of eukaryotic expression vector of lingual antimicrobial peptide(LAP) gene from mammary gland tissue of cow and its transient expression in COS-7 cells |
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| CAO Sui-zhong,YUAN Xi,LUAN Hong-yu,YU Shu-min,LIU Zong-ping.Construction of eukaryotic expression vector of lingual antimicrobial peptide(LAP) gene from mammary gland tissue of cow and its transient expression in COS-7 cells[J].Chinese Journal of Veterinary Science,2011,31(4). | |
| Authors: | CAO Sui-zhong YUAN Xi LUAN Hong-yu YU Shu-min LIU Zong-ping |
| Institution: | CAO Sui-zhong1,2,YUAN Xi1,LUAN Hong-yu1,YU Shu-min1,LIU Zong-ping2(1.College of Veterinary Medicine,Sichuan Agricultural University,Ya'an,Sichuan 625014,China,2.College of Veterinary Medicine,Yangzhou University,Yangzhou,Jiangsu 225009,China) |
| Abstract: | The lingual antimicrobial peptide(LAP) gene of dairy cattle was amplified from the mammary gland tissue suffered mastitis by RT-PCR,using a pair of special primers according to the sequence published in GenBank,and then cloned into the pMD19-T simple vector.The LAP gene was extracted by digesting the recombinant plasmid,and subcloned into the pEGFP-C1 vector to construct eukaryotic expression vector named as pEGFP-LAP.Finally,the constructed plasmid pEGFP-LAP was transfected into COS-7 cells by lipofectin t... |
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