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猪瘟病毒NS2 3′端基因的克隆与原核表达 被引量:2

Cloning and Prokaryotic Expression of NS2 Gene of CSFV
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摘要 从真核表达载体pEGFP-NS2中克隆得到猪瘟病毒(CSFV)NS2 3′端的基因片段,将其克隆到pET-32a载体上,构建出重组表达载体,经IPTG诱导表达。结果表明,克隆了大小为363 bp NS2 3′端的基因,重组载体pET-NS2-C363在大肠埃希菌中以包涵体形式表达,表达产物经SDS-PAGE和Western blot鉴定大小为33.84 ku,与预期结果一致。结果为进一步研究NS2蛋白与猪脐静脉血管内皮细胞中蛋白之间的相互作用提供材料及为制备单克隆抗体奠定基础。 The research studied cloning and expression of NS2 gene.363 bp of NS2 gene of CSFV Shimen strain was amplified by PCR,and cloned into prokaryotic expression vector pET-32a.The target protein was expressed in E.coli BL21(DE3) induced with IPTG,and detected by SDS-PAGE and Western blot.The results indicated that the 33.84 ku expressed protein was obtained.,It laid the foundation to study the interaction between theporcine vascular endothelia cell protein andNS2 protein and monoclonal antibody preparatio.
出处 《动物医学进展》 CSCD 北大核心 2011年第5期14-17,共4页 Progress In Veterinary Medicine
基金 国家自然科学基金项目(30972186)
关键词 猪瘟病毒 NS2基因 克隆 原核表达 Classical swine fever virus NS2 gene coling prokaryotic expression
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参考文献11

  • 1Franeki R, Fauguet C M, Knudson D, et al. Flaviviridae[J]. Arch Virol Suppl,1991,2: 223-233.
  • 2Lackner T, Thiel H J, Tautz N. Dissection of a viral autoprotease elucidates a function of a cellular chaperone in proteolysis [J]. Proc Natl Aead Sci U S A,2006,103(5): 1510-1515.
  • 3Tang Q H,Zhang Y M,Fan L,et al. Classic swine fever virus NS2 protein leads to the induction of cell cycle arrest at S-phase and endoplasmic reticulum stress[J]. J Virol,2010, 7: 4.
  • 4Tang Q H, Zhang Y M, Xu Y Z,et al. Up-regulation of integrin beta 3expression in porcine vascular endothelial cells cultured in vitro by classical swine fever virus[J]. Vet Immunol Immunopathol, 2010,113:237-242.
  • 5Agapov E V,Murray C L, Frolov I, et al. Uncleared NS2-3 is required for production of infectious bovine viral diarrhea virus [J]. J Virol,2004,78(5): 2414-2425.
  • 6Moulin H R, Seuberlieh T, Bauhofer O, et al. Nonstructural proteins NS2-3 and NS4A of classical swine fever virus: essential features for infectious particle formation [J]. Virology, 2007,365(2) : 376-389.
  • 7Lackner T, Thiel H J, Tautz N. Dissection of a viral autoprotease elucidates a funetion of a cellular chaperone in proteolysis [J]. Proc Natl Acad SeiUSA,2006,103(5): 1510-1515.
  • 8Peter K, Karl-Josef D. Identification of DNA-binding proteins and protein-protein interactions by yeast one-hybrid and yeast two-hybrid screen[J]. Methods Mol Biol,2010,639: 171-192.
  • 9孔建强,沈君豪,黄勇,阮仁余,项斌,郑晓东,程克棣,王伟.甲型H1N1流感病毒非结构蛋白NS1与人CPSF30结合拮抗剂筛选模型的建立及药物筛选[J].药学学报,2010,45(3):388-394. 被引量:5
  • 10Pajunen M, Poussu E, Turakainen H, et al. Application of Mu in vitro transposition for high-precision mapping of protein-protein interfaces on a yeast two-hybrid platform[J]. Methods,2009, 49(3) :255-262.

二级参考文献21

  • 1Bornholdt ZA, Prasad BV. X-ray structure of influenza virus NS1 effector domain [J]. Nat Struct Mol Biol, 2006, 13: 559-560.
  • 2Burgui I, Aragon T, Ortin J, et al. PABP1 and elF4GI associate with influenza virus NS 1 protein in viral mRNA translation initiation complexes [J]. J Gen Virol, 2003, 84: 3263-3274.
  • 3Delgadillo MO, Saenz P, Salvador B, et al. Human influenza virus NS1 protein enhances viral pathogenicity and acts as an RNA silencing suppressor in plants [J]. J Gen Virol, 2004, 85: 993-999.
  • 4Enami M, Enami K. Characterization of influenza virus NS1 protein by using a novel helper-virus-free reverse genetic system [J]. J Virol, 2000, 74: 5556-5561.
  • 5Bergmann M, Garcia-Sastre A, Carnero E, et al. Influenza virus NS1 protein counteracts PKR-mediated inhibition of replication [J]. J Virol, 2000, 74: 6203-6206.
  • 6Ehrhardt C, Wolff T, Pleschka S, et al. Influenza A virus NS 1 protein activates the PI3k/Akt pathway to mediate antiapoptofic signaling responses [J]. J Virol, 2007, 81: 3058-3067.
  • 7Hale BG, Jackson D, Chen YH, et al. Influenza A virus NS1 protein binds p85beta and activates phosphatidylinositol-3- kinase signaling [J]. Proc Natl Acad Sci USA, 2006, 103: 14194-14199.
  • 8Stasakova J, Ferko B, Kittel C, et al. Influenza A mutant viruses with altered NS1 protein function provoke caspase-1 activation in primary human macrophages, resulting in fast apoptosis and release of high levels of interleukins 1B and 18 [J]. J GenVirol, 2005, 86: 185-195.
  • 9Guo Z, Chen LM, Zeng H, et al. NS 1 protein of influenza A virus inhibits the function of intracytoplasmic pathogen sensor, RIG-I [J]. Am J Respir Cell Mol Biol, 2007, 36: 263-269.
  • 10Min JY, Krug RM. The primary function of RNA binding by the influenza A virus NS1 protein in infected cells: inhibiting the 2'-5' oligo (A) synthetase/RNase L pathway [J]. Proc Natl Acad Sci USA, 2006, 103: 7100-7105.

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