期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

Hepc20在毕赤酵母中的胞内表达及鉴定

Intracellular expression and identification of Hepc 20 in Pichia pastoris
下载PDF
导出
摘要 构建了Hepc 20的毕赤酵母表达载体,在毕赤酵母中能成功表达有活性的Hepc 20。优化了菌株的培养诱导条件,结果表明BMMY培养基是Hepc 20表达和重组菌株生长的最佳培养基,甲醇诱导终体积分数0.5%,在此条件下Hepc 20在重组菌株中的表达量约为3.6 mg/L。Western blot检测显示的22000处的条带为重组Hepc 20条带;ELISA验证表明重组Hepc 20可以跟抗体特异结合,琼脂扩散法抗菌实验表明重组Hepc 20对金黄色葡萄球菌和枯草芽孢杆菌表现出抗菌活性。 A recombinant Pichia pastoris strain for Hepc 20 production has been constructed and an active hepcidin peptide was successfully expressed in methylotrophic yeast. A buffered methanol complex （BMMY） medium was found to be optimal for the hepcidin protein expression and growth of the recombinant strains, and when the final volume fraction of methanol was 0.5% , the expression level of hepcidin protein reached the maximum value of 3.6 mg/L. Western blotting showed the 22000 band of the recombinant hepcidin protein whilst enzyme-linked immu- nosorbent （ELISA） assay showed that the recombinant Hepc 20 peptide can combine with specific antibodies, and the agar diffusion technique showed that Hepc 20 exhibited antibacterial activity against S. aureus and B. subtilis.
作者 Farzana RASHID 袁其朋 孙新晓 李飞
机构地区 北京化工大学生命科学与技术学院
出处 《北京化工大学学报（自然科学版）》 CAS CSCD 北大核心 2010年第2期104-108,共5页 Journal of Beijing University of Chemical Technology(Natural Science Edition)
基金 国家自然科学基金(20576010)
关键词 抗菌肽 Hepc 20 铁 毕赤酵母 antimicrobial peptide hepcidin 20 iron Pichia pastoris
分类号 Q786 [生物学—分子生物学]
  • 相关文献

参考文献10

  • 1Krause A, Neitz S, Magert H J, et al. LEAP-1, a novel highly disulfide bonded human peptide, exhibits antimicrobial activity[ J ]. FEBS Letters, 2000, 480 ( 2 ) : 147 -150.
  • 2Sima P, Trebichavsky, I, Sigler K. Mammalian antibiotic peptides[ J]. Folia Microbiol, 2003, 48(2) : 123 - 137.
  • 3Nemeth E, Rivera S, Gabayan V, et al. IL-6 mediates hypoferremia of inflammation by inducing the synthesis of the iron regulatory hormone hepcidin[ J]. Journal of Clinical Investigation, 2004, 113 ( 9 ) : 1271 - 1276.
  • 4Zhang L, Falla T, Wu M, et al. Determinants of recombinant production of antimicrobial cationic peptides and creation of peptide variants in bacteria [ J ]. Biochemical and Biophysical Research Communications, 1998, 247 (3) : 674 - 680.
  • 5Park C H, Valore E V, Waring A J, et al. Hepcidin, a urinary antimicrobial peptide synthesized in the liver[ J]. The Journal of Biological Chemistry, 2001, 276 ( 11 ) : 7806 - 7810.
  • 6Koliaraki V, Marinou M, Samiotaki M, et al. Iron regulatory and bactericidal properties of human recombinant hepcidin expressed in Pichia pastoris [ J ]. Biochimie, 2008, 90(5) : 1 -10.
  • 7Schaggar H. Trince-SDS-PAGE [ J]. Nature Protocols, 2006, 1(1): 16-20.
  • 8Koganesawa N, Aizawa T, Shimojo H, et al. Expression and purification of a small cytokine growth-blocking peptide from armyworm Pseudaletia separata by an optimized fermentation method using the methylotrophic yeast Pichia pastoris[ J ]. Protein Expression and Purification, 2002, 25(3) : 416 -425.
  • 9Ru-Shi Liu Kun-Yu Yang Jian Lin Yi-Wei Lin Zhi-Hong Zhang Jun Zhang Ning-Shao Xia The Key Laboratory for Cell Biology and Tumor Cell Engineering of the Ministry of Education,The Research Center for Medical Molecular Virology of Fujian Province,Xiamen University,Xiamen 361005,Fujian Province,China.High-yield expression of recombinant SARS coronavirus nudeocapsid protein in methylotrophic yeast Pichia pastoris[J].World Journal of Gastroenterology,2004,10(24):3602-3607. 被引量：8
  • 10Boettner M, Prinz B, Holz C, et al. High-throughput screening for expression of heterologous proteins in the yeast Pichia pastoris [ J ]. Journal of Biotechnology, 2002, 99 ( 1 ) : 51 - 60.

二级参考文献30

  • 1[1]Poutanen SM, Low DE, Henry B, Finkelstein S, Rose D, Green K,Tellier R, Draker R, Adachi D, Ayers M, Chan AK, Skowronski DM, Salit I, Simor AE, Slutsky AS, Doyle PW, Krajden M,Petric M, Brunham RC, McGeer AJ. Identification of severe acute respiratory syndrome in Canada, N Engl J Med 2003;348:1995-2005
  • 2[2]Lee N, Hui D, Wu A, Chan P, Cameron P, Joynt GM, Ahuja A,Yung MY, Leung CB, To KF, Lui SF, Szeto CC, Chung S, Sung JJ. A major outbreak of severe acute respiratory syndrome in Hong Kong. N Engl J Med 2003; 348:1986-1994
  • 3[3]Peiris JS, Lai S, Poon L, Guan Y, Yam LY, Lim W, Nicholls J,Yee WK, Yan WW, Cheung MT, Cheng VC, Chan KH, Tsang DN, Yung RW, Ng TK, Yuen KY. Coronavirus as a possible cause of severe acute respiratory syndrome. Lancet 2003;361:1319-1325
  • 4[4]Ksiazek TG, Erdman D, Goldsmith CS, Zaki SR, Peret T, Emery S, Tong S, Urbani C, Comer JA, Lim W, Rollin PE, Dowell SF,Ling AE, Humphrey CD, Shieh WJ, Guarner J, Paddock CD,Rota P, Fields B, DeRisi J, Yang JY, Cox N, Hughes JM, LeDuc JW, Bellini WJ, Anderson LJ. A novel coronavirus associated with severe acute respiratory syndrome. N Engl J Med 2003;348:1953-1966
  • 5[5]Drosten C, Gunther S, Preiser W, van der Werf S, Brodt HR,Becker S, Rabenau H, Panning M, Kolesnikova L, Fouchier RA,Berger A, Burguiere AM, Cinatl J, Eickmann M, Escriou N,Grywna K, Kramme S, Manuguerra JC, Muller S, Rickerts V,Sturmer M, Vieth S, Klenk HD, Osterhaus AD, Schmitz H,Doerr HM. Identification of a novel coronavirus in patients with severe acute respiratory syndrome. N Engl J Med 2003;348:1967-1976
  • 6[6]Anand K, Ziebuhr J, Wadhwani P, Mesters JR, Hilgenfeld R.Coronavirus main mroteinase (3CLpro) structure: basis for design of anti-SARS drugs. Science 2003; 300:11763-11767
  • 7[7]Marra MA, Jones SJM, Astell CR, Holt RA, Brooks-Wilson A,Butterfield YSN, Khattra J, Asano JK, Barber SA, Chan SY,Cloutier A, Coughlin SM, Freeman D, Girn N, Griffith OL,Leach SR, Mayo M, McDonald H, Montgomery SB, Pandoh PK, Petrescu AS, Robertson AG, Schein JE, Siddiqui A, Smailus DE, Stott JM, Yang GS, Plummer F, Andonov A, Artsob H,Bastien N, Bernard K, Booth TF, Bowness D, Czub M, Drebot M, Fernando L, Flick R, Garbutt M, Gray M, Grolla A, Jones S,Feldmann H, Meyers A, Kabani A, Li Y, Normand S, Stroher U,Tipples GA, Tyler S, Vogrig R, Ward D, Watson B, Brunham RC, Krajden M, Petric M, Skowronski DM, Upton C, Roper RL.The genome sequence of the SARS-associated coronavirus. Science 2003; 300:1399-1345
  • 8[8]Rota PA, Oberste MS, Monroe SS, Nix WA, Campagnoli R,Lcenogle JP, Penaranda S, Bankamp B, Maher K, Chen MH,Tong S, Tamin A, Lowe L, Frace M, DeRisi JL, Chen Q, Wang D, Erdman DD, Peret TCT, Burns C, Ksiazek TG, Rollin PE,Sanchez A, Liffick S, Holloway B, Limor J, McCaustland K,Olsen-Rasmussen M, Fouchier R, Gunther S, Osterhaus ADME,Drosten C, Pallansch MA, Anderson LJ, Bellini WJ. Characterization of a novel coronavirus associated with severe acute respiratory syndrome. Science 2003; 300:1394-1399
  • 9[9]Gomez N, Carrillo C, Salinas J, Parra F, Borca MV, Escribano JM. Expression of immunogenic glycoprotein S polypeptides from transmissible gastroenteritis coronavirus in transgenic plants. Virology 1998; 249:352-358
  • 10[10]Jackwood MW, Hilt DA. Production and immunogenicity of multiple antigenic peptide (MAP) constructs derived from the S1 glycoprotein of infectious bronchitis virus (IBV). Adv Exp Med Biol 1995; 308:213-219

共引文献7

北京化工大学学报（自然科学版）
2010年 第2期

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部