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高致病性猪繁殖与呼吸综合征病毒蛋白质组双向电泳方法的建立及优化 被引量:2

Establishment and optimization of two-dimensional electrophoresis for proteome of highly pathogenic porcine reproductive and respiratory syndrome virus
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摘要 为建立高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)蛋白质组双向电泳方法,本研究通过对HP-PRRSV蛋白样品提取、裂解、一向等电聚程序、上样量、染色试剂、显色时间等条件优化,建立了有效的HP-PRRSV蛋白质组学双向电泳方法。优化结果表明采用冻融-超声-裂解提取样品,结合2-D clean-up试剂盒纯化蛋白,按上样量为200μg,采用硝酸银染色,显色6 min,选择适宜的一向等电聚焦参数,能获得分辨率高、重复性好的双向电泳图谱。HP-PRRSV蛋白质组双向电泳方法的建立,为开展该病毒蛋白质组和免疫蛋白质组研究奠定了基础。 To establish an efficient 2-DE protocol for proteomic method of highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV), the conditions were compared and optimized, including the sample preparation, IEF, appropriate strips, staining reagent and treatment. The results showed that the high resolving and repeatable 2-DE images were obtained with 200 ~g of PRRSV protein samples which was treated consecutively with lysate buffer, frozen-thaw for four times, sonicated for 5 min, purified by 2-D Clean-up kit, and subjected to 2-DE separation under suitable electrophoresis parameters and visualized by silver-staining for 6 min. This study would facilitate future study on proteome and immunoprotein of highly pathogenic PRRSV.
出处 《中国预防兽医学报》 CAS CSCD 北大核心 2012年第5期365-369,共5页 Chinese Journal of Preventive Veterinary Medicine
基金 福建省杰出青年科学基金项目(2009J06012)
关键词 高致病性猪繁殖与呼吸综合征病毒 蛋白质组 双向电泳 highly pathogenic PRRSV proteome two-dimensional electrophoresis
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  • 1Tian Ke-gong, Yu Xiu-ling, Zhao Tie-zhu, et al. Emergence of fatal PRRSV variants: unparalleled outbreaks of atypical PRRS in China and molecular dissection of the unique hallmark [J]. PLoS One, 2007, 2(6): e526.
  • 2De Lima M, Pattnaik A K, Flores E F, et al. Serologic marker candidates identified among B-cell linear epitopes of NSP2 and structural proteins of a North Ameriean sWain of porcine repro- ductive and respiratory syndrome virus [J]. Virology, 2006, 353 (2): 410-421.
  • 3童光志,周艳君,郝晓芳,田志军,仇华吉,彭金美,安同庆.高致病性猪繁殖与呼吸综合征病毒的分离鉴定及其分子流行病学分析[J].中国预防兽医学报,2007,29(5):323-327. 被引量:390
  • 4周伦江,王隆柏,王伟,魏宏,车勇良,陈如敬,庄向生.变异型猪繁殖与呼吸综合征病毒FJ06A毒株的分离鉴定和致病性[J].福建农林大学学报(自然科学版),2011,40(6):618-623. 被引量:4
  • 5Wilkins M R, Sanchez J C, Gooley A A, et al. Progress with proteome p rojects: why all proteins expressed by a genome should be identified and how to do it [J]. Biotechnol Genet Eng Rev, 1996, 13: 19-50.
  • 6Chelius D, Htihmer A F, Shieh C H, et al. Analysis of the aden- ovirus type 5 proteome by liquid chromatography and tandem mass spectrometry methods [J]. J Proteome Res, 2002, 1 (6): 501-513.
  • 7Chertova E, Chertov O, Coren L V, et al. Proteomic and bio- chemical analysis of purified human immunodeficiency virus type 1 produced from infectedmonocyte-derived macrophages [J]. J Virol, 2006, 80(18): 9039-9052.
  • 8张爱玲,王兴龙,王英超,党源,张复贤.2型猪链球菌分泌组双向电泳样品制备方法的建立[J].中国预防兽医学报,2009,31(10):776-779. 被引量:6
  • 9张娜娜,许勤,李风良,程罗根.小菜蛾成虫蛋白质组双向电泳图谱的建立及条件优化[J].南京师大学报(自然科学版),2011,34(2):78-82. 被引量:4
  • 10Nagalla S R, Canick J A, Jacob T, et al. Proteomic analysis of maternal serum in down syndrome: identification of novel pro- tein biomarkers [J]. J Proteome Res, 2007, 6(4): 1245-1257.

二级参考文献54

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同被引文献22

  • 1王晨燕,王隆柏,车勇良,吴学敏,陈如敬,邵良平,周伦江.应用双向电泳分析2株不同毒力的副猪嗜血杆菌蛋白质表达的差异[J].畜牧兽医学报,2015,46(3):503-508. 被引量:1
  • 2乐军,蒋晓飞,梁莉,吕元.蛋白质组学分析多重耐药的铜绿假单胞菌[J].中国抗生素杂志,2006,31(8):496-500. 被引量:4
  • 3斯特劳,刘文军,张中秋,等.猪病学[M].第8版.北京:中国农业大学出版社,1998.
  • 4Simone Oliveira,Carlos Pijoan.Haemophilus parasuis : new trends on diagnosis, epidemiology and control[J]. Veterinary Microbiology . 2003 (1)
  • 5Virginia Aragon,Marta Cerdà-Cuéllar,Lorenzo Fraile,Mark Mombarg,Miquel Nofrarías,Alexandre Olvera,Marina Sibila,David Solanes,Joaquim Segalés.Correlation between clinico-pathological outcome and typing of Haemophilus parasuis field strains[J]. Veterinary Microbiology . 2009 (3)
  • 6Gisele G. Tomazella,Kine Risberg,Haima Mylvaganam,Paul Christoffer Lindemann,Bernd Thiede,Gustavo A. de Souza,Harald G. Wiker.Proteomic analysis of a multi-resistant clinical Escherichia coli isolate of unknown genomic background[J]. Journal of Proteomics . 2011 (6)
  • 7Bin Zhang,Chenggang Xu,Suming Zhou,Saixiang Feng,Lingyun Zhang,Yanbin He,Ming Liao.Comparative proteomic analysis of a Haemophilus parasuis SC096 mutant deficient in the outer membrane protein P5[J]. Microbial Pathogenesis . 2011 (2)
  • 8Deb N Chakravarti,Michael J Fiske,Leah D Fletcher,Robert J Zagursky.Application of genomics and proteomics for identification of bacterial gene products as potential vaccine candidates[J]. Vaccine . 2001 (6)
  • 9Malinverni Juliana C,Werner John,Kim Seokhee,Sklar Joseph G,Kahne Daniel,Misra Rajeev,Silhavy Thomas J.YfiO stabilizes the YaeT complex and is essential for outer membrane protein assembly in Escherichia coli. Molecular Microbiology . 2006
  • 10Olvera A,Cerdà-Cuéllar M,ARAGON V.Study of the popu-lation structure of Haemophilus parasuis by multilocus se-quence typing. Microbiology . 2006

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