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链球菌表面烯醇化酶及其变异体重组蛋白的表达纯化 被引量：1

EXPRESSION AND PURIFICATION OF RECOMBINANT STREPTOCOCCAL SURFACE ENOLASE AND C-TERMINAL LYSINE RESIDUES-TRUNCATED VARIANT
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摘要 目的:表达并纯化M6型GAS表面烯醇化酶以及其敲除C末端赖氨酸残基重组蛋白(rSEN和rSENΔ434-435)。方法:克隆了M6型GAS ATCC32175的SEN基因以及SENΔ434-435基因,与pASK-IBA37载体连接后,表达蛋白并用亲和层析色谱纯化重组蛋白;通过酶促反应实验测定了重组蛋白的生物活性。结果:2种基因克隆条件稳定,蛋白表达量大,且纯化方法可靠,纯化的重组蛋白具有较高的生物活性。结论:成功表达并纯化了rSEN和rSENΔ434-435蛋白。 Objective：To express and purify the recombinant streptococcal surface enolase （rSEN） and its C -terminal lysine residues -truncated variant （rSENA434 -435）. Methods：We cloned sen and sen△434 -435 from M6 -type GAS ATCC32175, produced rSEN and rSEN△434 - 435 in E. coli using the 6 × Histag pASK - IBA37 expression vector and purified the recombinant proteins by af- finity chromatography with TALON metal affinity resins. The enzyme reaction was performed to determine enolase activity. Results： PCR conditions ampifing sen and sen△434 -435 were veridical and expression and purification of recombinant proteins were profuse and convenient. The purified rSEN and rSEN△434 - 435 were found to have relatively full enolase activity. Conclusion ： We succusfully expressed and purified rSEN and rSEN△434 -435.
作者 许丽萍 白文成 刘杨 代霄燕 李培峰
机构地区 内蒙古农业大学兽医学院 内蒙古农业大学生命科学学院
出处 《内蒙古农业大学学报（自然科学版）》 CAS 北大核心 2011年第1期167-171,共5页 Journal of Inner Mongolia Agricultural University(Natural Science Edition)
基金 国家自然科学基金(30860019)
关键词 A群链球菌 烯醇化酶 表达 纯化 Group A streptococcus α - enolase expression purification
分类号 Q936 [生物学—微生物学]
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参考文献13

  • 1Cunningham MW. Pathogenesis of group A streptococcal infections[J]. Clin Microbiol Rev, 2000, 13 (3) : 470 -511.
  • 2Cunningham MW. Pathogenesis of group A streptococcal infections and their sequelae [ J ]. Adv Exp Med Biol, 2008, 609 29 - 42.
  • 3Sun H, Ringdahl U, Homeister JW, et al. Plasminogen is a critical host pathogenicity factor for group A streptococcal infection[J]. Science, 2004, 305 (5688) : 1283 - 1286.
  • 4Walker M J, McArthur JD, McKay F, et al. Is plasminogen deployed as a Streptococcus pyogenes virulence factor? [J]. Trends Microbiol, 2005, 13 (7) : 308 -313.
  • 5Berge A, Sjobring U. PAM, a novel plasminogen -binding protein from Streptococcus pyogenes [ J ]. J Biol Chem, 1993, 268 (34): 25417-25424.
  • 6Svensson MD, Sjobrlng U, Bessen DE. Selective distribution of a high - affinity plasminogen - binding site among group A streptococci associated with impetigo [ J]. Infect Immun, 1999, 67 (8): 3915-3920.
  • 7Derbise A, Song YP, Parikh S, et al. Role of the C -terminal lysine residues of streptococcal surface enolase in Glu - and Lys - plasminogen - binding activities of group A streptococci [ J ].Infect Immun, 2004, 72 ( 1 ) : 94 - 105.
  • 8Ranson M, Andronicos NM. Plasminogen binding and cancer: promises and piffalls[J]. Front Biosci, 2003, 8 s294 - 304.
  • 9Cork A J, Jergie S, Hammersehmidt S, et al. Defining the structural basis of human plasminogen binding by streptococcal surface enolase [ J ]. J Biol Chem, 2009, 284 (25) : 17129 - 17137.
  • 10Pancholi V, Fischetti VA. alpha- enolase, a novel strong plasmin(ogen) binding protein on the surface of pathogenic streptococci [ J]. J Biol Chem, 1998, 273 (23) : 14503 - 14515.

同被引文献8

  • 1Turk D. The pathogenicity of HaemopH. influenzalus influ- enzae[J]. Journal of medical microbiology, 1984,18 : 1 - 16.
  • 2Purification and characterisation of a plasminogen- binding protein from Haemophilus influenzae. Sequence determina- tion reveals identity with aspartase [J]. Biochimica et Bio- physica Acta, 1997,1324 : 182 - 190.
  • 3Diana Barthel, Birendra Singh, Kristian Riesbeck, et al. Heamophilus influenczeae Uses the Surface Protein E To Acquire Human Plasminogen and To Evade Innate Immunity [J]. J Immunol, 2012,188:379 - 385.
  • 4Lottenberg R, Broder CC, Boyle MD, et al. Cloning, se-quence analysis, and expression in EschericH. influenzaa coli of a streptococcal plasmin receptor [J].J BacterioI, 1992, 174 (16): 5204-5210.
  • 5McLean JW, Tomlinson JE, Kuang WJ, et al. cDNA se- quence of human apolipoprotein (a) is homologous to plas- minogen[J], Nature. 1987,330:132-137.
  • 6Rocco Romagnuolo, Santica M. Marcovina, Michael B. Boffa, et al. Inhibition of plasminogen activation by apo (a) : role of carboxyl - terminal lysines and identification of inhibitory domains in apo(a) [J]. J Lipid Res. 2014 Apr; 55 (4) : 625 - 634.
  • 7Han R. Plasma lipoproteins are important components of the immune system[J]. Microbiol Immunol. 2010 : 54 : 246 - 253.
  • 8Williams VR and DJ Lartigue. Quaternary structure and certain allosteric properties of aspartase[J]. Journal of Bio- logical Chemistry, 1967. 242(12) : 2973 -2978.

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内蒙古农业大学学报（自然科学版）
2011年 第1期

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