[1]郭长明,袁橙,朱善元,等.应用iTRAQ定量蛋白质组学技术筛选无乳链球菌鱼源株与牛源株差异表达蛋白[J].江苏农业学报,2017,(04):868-873.[doi:doi:10.3969/j.issn.1000-4440.2017.04.022]
 GUO Chang-ming,YUAN Cheng,ZHU Shan-yuan,et al.Quantitative identification of differential proteins in Streptococcus agalactiae piscine strain and bovine strain using iTRAQ[J].,2017,(04):868-873.[doi:doi:10.3969/j.issn.1000-4440.2017.04.022]
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应用iTRAQ定量蛋白质组学技术筛选无乳链球菌鱼源株与牛源株差异表达蛋白()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2017年04期
页码:
868-873
栏目:
畜牧兽医·水产养殖
出版日期:
2017-08-30

文章信息/Info

Title:
Quantitative identification of differential proteins in Streptococcus agalactiae piscine strain and bovine strain using iTRAQ
作者:
郭长明1袁橙1朱善元1刘广锦2陆承平2刘永杰2
(1.江苏农牧科技职业学院/江苏省兽用生物制药高技术研究重点实验室,江苏泰州225300;2.南京农业大学动物医学院,江苏南京210095)
Author(s):
GUO Chang-ming1YUAN Cheng1ZHU Shan-yuan1LIU Guang-jin2LU Cheng-ping2LIU Yong-jie2
(1.Jiangsu Provincial Veterinary Bio-pharmaceutical High-tech Key Laboratory, Jiangsu Agri-animal Husbandry Vocational College, Taizhou 225300, China;2.College of Veterinary Medicine, Nanjing Agricultural University, Nanjing210095, China)
关键词:
无乳链球菌鱼源株牛源株iTRAQcpsIVK基因蛋白质组学
Keywords:
Stretococcus galactiaepiscine strainbovine strainiTRAQcpsIVK geneproteomics
分类号:
S941.42
DOI:
doi:10.3969/j.issn.1000-4440.2017.04.022
文献标志码:
A
摘要:
为揭示不同宿主来源无乳链球菌致病机制,提取无乳链球菌鱼源株GD201008-001和牛源株ATCC13813的全菌蛋白质,经iTRAQ试剂标记后进行质谱鉴定,质谱数据用软件Mascot2.2和ProteomeDiscoverer14进行查库(UniProt数据库)鉴定及定量分析,并利用blast2go软件对差异蛋白质进行GO(gene ontology)功能注释。共鉴定出在鱼源株GD201008-001和牛源株ATCC13813中差异表达的蛋白质17个(P<005),其中在ATCC13813中上调表达的蛋白质3个(比值>1500),下调表达的蛋白质14个(比值<0667)。GO分析预测结果表明这些蛋白质主要参与15个生物学功能,涉及代谢过程、细胞成分、结合、催化活性、结构分子活性等。其中cpsIVK在牛源株ATC13813中下调最显著,其功能为荚膜生物合成蛋白质,与无乳链球菌荚膜合成相关。进一步验证发现,鱼源株GD201008-001抵抗小鼠巨噬细胞RAW264.7吞噬的能力显著高于cpsIVK下调的牛源株ATCC13813(P<001),提示cpsIVK可能在鱼源无乳链球菌逃避宿主免疫细胞吞噬过程中发挥功能。
Abstract:
To identify differentially expressed proteins in Streptococcus agalactiae (S. galactiae) piscine strain (GD201008-001) and bovine strain (ATCC13813) by iTRAQ, and provide new clues for exploring the pathogenic mechanism of S. galactiaestrains isolated from different hosts, cellular proteins were extracted from cultures of S. galactiae GD201008-001 and ATCC13813, and labeled with isobaric tags for relative and absolute quantitation (iTRAQ). Differentially expressed proteins were identified with LC-MS/MS. The mass spectrometry data was analyzed by Mascot2.2 and Proteome Discoverer1.4 and subjected to biological information analysis by blast2go. A total of 17 differentially expressed proteins were revealed in GD201008-001 and ATCC13813 (P<005), among which 3 proteins were up-regulated (ratio>1.5) and 14 were down-regulated (ratio<0.667) in ATCC13813. These proteins are involed in 15 biological functions including metabolic process, cell composition, binding, catalytic activity, and hypothetical proteins. The most down-regulated protein cpsIVK encoding capsule biosynthesis protein was related to S. agalactiae capsular synthesis. Further experiments showed that the antiphagocytosis of GD201008-001 resisting murine macrophages RAW264.7 was significantly higher than ATCC13813 (P<001) in which cpsIVK was down-regulated, suggesting that cpsIVK may play a role in the resistance to host immune cell phagocytosis of S. galactiae piscine strain.

参考文献/References:

[1]KEEFE G P. Streptococcus agalactiae mastitis: a review[J]. Canadian Veterinary Journal La Revue Veterinaire Canadienne, 1997, 38(7):429.
[2]SUANYUK N, KONG F, KO D, et al. Occurrence of rare genotypes of Streptococcus agalactiae, in cultured red tilapia Oreochromis sp. and Nile tilapia O. niloticus, in Thailand-Relationship to human isolates[J]. Aquaculture, 2008, 284(1-4):35-40.
[3]刘溪,敖秋桅,谭芸, 等. 无乳链球菌感染吉富罗非鱼后体内病原菌的动态分布[J]. 南方农业学报, 2015,46(9):1715-1719.
[4]郭玉娟,张德锋,樊海平,等.中国南方地区罗非鱼无乳链球菌的分子流行病学研究[J].水产学报, 2012, 36(3):399-406.
[5]ROSS P L, HUANG Y N, MARCHESE J N, et al. Multiplexed protein quantitation in Saccharomyces cerevisiae using amine-reactive isobaric tagging reagents[J]. Molecular & Cellular Proteomics, 2004, 3(12):1154-1169.
[6]WU W W, WANG G, BAEK S J, et al. Comparative study of three proteomic quantitative methods, DIGE, cICAT, and iTRAQ, using 2D gel- or LC-MALDI TOF/TOF[J]. Journal of Proteome Research, 2006, 5(3):651-658.
[7]WISNIEWSKI J, ZOUGMAN A, NAGARAJ N, et al. Universal sample preparation method for proteome analysis[J]. Nature Methods, 2009, 6(5):359-362.
[8]SANDBERG A, LINDELL G, KLLSTRM B N, et al. Tumor proteomics by multivariate analysis on individual pathway data for characterization of vulvar cancer phenotypes[J]. Molecular & Cellular Proteomics Mcp, 2012, 11(7): 3929-3936.
[9]LUN S, WILLSON P J. Expression of green fluorescent protein and its application in pathogenesis studies of serotype 2 Streptococcus suis[J]. Journal of Microbiological Methods, 2004, 56(3):401-412.
[10]UNWIN R D, GRIFFITHS J R, WHETTON A D. Simultaneous analysis of relative protein expression levels across multiple samples using iTRAQ isobaric tags with 2D nano LC-MS/MS[J]. Nature Protocols, 2010, 5(9):1574-1582.
[11]袁建丰,李林林,孙敏华,等. iTRAQ标记技术及其在微生物比较蛋白质组学中的研究进展[J].中国预防兽医学报, 2013, 35(10):859-862.
[12]DORAN K, NIZET V. Molecular pathogenesis of neonatal group B streptococcal infection: no longer in its infancy[J]. Molecular Microbiology, 2004, 54(1):23-31.
[13]SPRATT B G, CROMIE K D. Penicillin-binding proteins of gram-negative bacteria[J]. Clinical Infectious Diseases, 1988, 10(4):699.
[14]GEORGOPAPADAKOU N H, LIU F Y. Penicillin-binding proteins in bacteria[J]. Antimicrobial Agents & Chemotherapy, 1980, 18(1):148-157.
[15]ZAPUN A, CONTRERASMARTEL C, VERNET T. Penicillin-binding proteins and beta-lactam resistance[J]. Fems Microbiology Reviews, 2008, 32(2):361-385.
[16]JONES A L, NEEDHAM R H, CLANCY A, et al. Penicillin-binding proteins in Streptococcus agalactiae: a novel mechanism for evasion of immune clearance[J]. Molecular Microbiology, 2003, 47(1):247-256.
[17]JONES A L, MERTZ R H, CARL D J, et al. A streptococcal penicillin-binding protein is critical for resisting innate airway defenses in the neonatal lung[J]. Journal of Immunology, 2007, 179(5):3196-3202.
[18]郭长明. 鱼源无乳链球菌感染小鼠巨噬细胞差异转录基因的筛选及功能研究[D]. 南京:南京农业大学, 2014.

备注/Memo

备注/Memo:
收稿日期:2017-01-22 基金项目:国家自然科学基金青年科学基金项目(31502085);江苏省自然科学基金青年科学基金项目(BK20140703);中央高校基本科研基金项目(KJQN201618);江苏农牧科技职业学院科研项目(NSF201503) 作者简介:郭长明(1984-),男,山东临朐人,博士,讲师,主要从事预防兽医学(病原微生物和寄生虫分子免疫方向)研究。(E-mail)gcmscience@126.com 通讯作者:刘永杰,(E-mail)liuyongjie@njau.edu.cn
更新日期/Last Update: 2017-09-01