| 真核表达载体pCDNA3-CaBP-D28k的构建及细胞表达 |
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| 引用本文： | 章世元,俞路,王雅倩,李燕舞,李治学,严桂芹,周联高.真核表达载体pCDNA3-CaBP-D28k的构建及细胞表达[J].西北农业学报,2008,17(5):11-15. |
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| 作者姓名： | 章世元 俞路 王雅倩 李燕舞 李治学 严桂芹 周联高 |
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| 作者单位： | 扬州大学动物科学与技术学院,扬州,225009 |
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| 摘 要： | 构建钙结合蛋白D28k(calbindin-D28k,CaBP-D28k)真核表达载体,并证实了其在真核细胞COS-1中的表达情况,为进一步研究该重组载体在动物体内的表达奠定了基础。通过酶切方法自含鸡CaBP-D28k基因的克隆质粒pGEM-T-CaBP-D28k中扩增CaBP-D28k基因,并将其克隆到真核表达载体pCDNA3中,阳性克隆鉴定后,在PEI作用下转染COS-1细胞,通过间接免疫荧光试验(IFA)检测到了CaBP-D28k在COS-1中的表达。
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| 关 键 词： | Calbindin-D28k 转染 IFA |
| 收稿时间： | 2008/1/8 0:00:00 |
| 修稿时间： | 2008/3/15 0:00:00 |
Construction of Eukaryotic Expressing Plasmid of Chicken CaBP-D28k Gene and Its Expression in Cell |
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| ZHANG Shi-yuan,YU Lu,WANG Ya-qian,LI Yan-wu,LI Zhi-xue,YAN Gui-qin and ZHOU Lian-gao.Construction of Eukaryotic Expressing Plasmid of Chicken CaBP-D28k Gene and Its Expression in Cell[J].Acta Agriculturae Boreali-occidentalis Sinica,2008,17(5):11-15. |
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| Authors: | ZHANG Shi-yuan YU Lu WANG Ya-qian LI Yan-wu LI Zhi-xue YAN Gui-qin and ZHOU Lian-gao |
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| Institution: | College of Animal Science and Technology, Yangzhou University, Yangzhou Jiangsu 225009, China;College of Animal Science and Technology, Yangzhou University, Yangzhou Jiangsu 225009, China;College of Animal Science and Technology, Yangzhou University, Yangzhou Jiangsu 225009, China;College of Animal Science and Technology, Yangzhou University, Yangzhou Jiangsu 225009, China;College of Animal Science and Technology, Yangzhou University, Yangzhou Jiangsu 225009, China;College of Animal Science and Technology, Yangzhou University, Yangzhou Jiangsu 225009, China;College of Animal Science and Technology, Yangzhou University, Yangzhou Jiangsu 225009, China |
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| Abstract: | Calbindin-D28k eukaryotic expression bearer was constructed and the expression in eucaryotic cell COS-1 was confirmed in this article,which could establish foundation for further researching the expression of recombined bearer.Chicken CaBP-D28k gene was amplified from the recombinant plasmid named pGEM-T-CaBP-D28k by digested(Xho I Bam HI) and cloned into pCDNA3.The recombinant plasmid was transfected into COS-1 with PEI.72 hours after transfection,the expression of CaBP-D28k were successfully tested by indirect immunofluorescence assay(IFA) with the antiserum of CaBP-D28k expressed in COS-1. |
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| Keywords: | Calbindin-D28k Transfection IFA |
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