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人胎盘膜联蛋白V cDNA的克隆及检测 被引量：1

cDNA CLONING AND DETERMINATION OF HUMAN PLACENTA ANNEXIN V GENE
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摘要 提取新鲜人胎盘组织的总RNA,利用RT-PCR技术扩增出膜联蛋白V(Annexin V)基因的编码序列,将扩增产物克隆到真核表达载体pCMV5并转化到大肠杆菌DH5α中.对重组质粒进行DNA测序,其测序结果完全正确.成功地从人胎盘组织中克隆出Annexin V的全长基因并构建了该基因的真核表达载体,为今后该基因的表达及其生理作用的研究奠定了基础. Annexin Ⅴ gene containing EcoR Ⅰ and Hind Ⅲ endoenzyme site is produced by using RT-PCR technique from fresh human placentas. The Annexin Ⅴ gene is digested by double enzyme （EcoR Ⅰ and Hind Ⅲ） and inserted into expression vector pCMV5. The ligation products are transformed into E. coli DH5α. Recombinant plasmid is determined by restriction endonuclease digestion and PCR methods. The result is in agreement with the expected data and DNA sequencing is carried out also. The Annexin Ⅴ gene is cloned and the recombinant eukaryotic expression plasmid containing human Annexin Ⅴ gene is successfully constructed. The production and functional assay of human Annexin Ⅴ protein are in progress.
作者 张鑫蕊 井健
机构地区 北京师范大学生命科学学院
出处 《北京师范大学学报（自然科学版）》 CAS CSCD 北大核心 2007年第4期438-441,共4页 Journal of Beijing Normal University(Natural Science)
基金 国家自然科学基金资助项目(30540071)
关键词 膜联蛋白V RT-PCR 真核表达载体 抗凝血 Annexin Ⅴ RT-PCR eukaryotic expression plasmid anticoagulant activity
分类号 Q78 [生物学—分子生物学]
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参考文献11

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同被引文献10

  • 1Gerke V, Moss S E. Annexins: from structure to function [J]. Physiol Rev, 2002, 82(2):331.
  • 2Pollard H B, haigler H T. Calcium-activated endonexin Ⅱ form calcium channels across acidic phospholipid bilayer membranes [J]. Biol Chem, 1990, 265: 1207.
  • 3Brisson A, Mosser G, Huber R. Structure of soluble and membrane-bound human annexin Ⅴ [ J ]. J Mol Biol, 1991, 220(2):199.
  • 4Funakoshi T, Heimark R L, Hendrickson L E, et al. Human placental anticoagulant protion: isolation and characterization [J]. Biochemistry, 1987, 26(17) :5572.
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  • 8Eberhard Neumann U, Peter M, Siemens Katja Toensing. Electroporative fast pore-flickering of the annexin Ⅴ-lipid surface complex, a novel gating concept for ion transport [J]. Biophysical Chemistry, 2000, 86 : 203.
  • 9Rand J H. Antiphospholipid antibody syndrome: new insight on thrombogenic mechanisms [J]. Am J Med Sci, 1998,2:142.
  • 10Vermes L Haanen C, Steffensnakken H, et al. A novel assay for poptosis: flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluoreseein labeled annexinv[J]. Journal Immunol Methods, 1995,184:39.

引证文献1

北京师范大学学报（自然科学版）
2007年 第4期

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