基于iTRAQ方法分析副溶血弧菌活的非可培养状态的差异表达上调蛋白.pdfVIP

现代食品科技 Modern Food Science and Technology 2015, Vol.31, No.8 基于 iTRAQ方法分析副溶血弧菌活的非可培养状态的 差异表达上调蛋白 田娟，叶忱，刘羽霏，王丽，钟青萍 （华南农业大学食品学院，广东广州 510642 ） 摘要：采用低温、寡营养和食品防腐剂处理诱导副溶血弧菌进入活的非可培养状态（viable but nonculturable state, VBNC ），并利 用同位素标记相对与绝对定量(isobaric rags for relative and absolute quantitation ，iTRAQ)蛋白组学技术分析处于VBNC 状态和对数生长 期的副溶血弧菌的差异表达蛋白，结合液质联用技术对蛋白进行鉴定，对差异蛋白进行GO 富集分析和Pathway 通路分析。结果表明， 副溶血弧菌于山梨酸钾浓度为10 mmol/L 的3% NaCl 寡营养培养基中，4 ℃条件下40~45 d 即可进入到VBNC 状态。通过iTRAQ 蛋 白组学技术共鉴定到135880 个蛋白，其中定量的蛋白有1088 个；在VBNC 状态显著下调蛋白36 个，上调蛋白15 个。差异表达的 上调蛋白主要是外膜蛋白、转运蛋白、铁蛋白和其他参与代谢的关键蛋白，表明 VBNC 状态的副溶血弧菌根据环境应激提高这些蛋 白的表达以保持活性。该研究结果为更深入探讨VBNC 的分子形成机制提供了理论基础。 关键词：副溶血弧菌；iTRAQ；活的非可培养状态；差异蛋白 文章篇号：1673-9078(2015)8-53-58 DOI: 10.13982/j.mfst.1673-9078.2015.8.010 The Up-regulated Expressed Proteins of the Viable but Nonculturable State of Vibrio parahaemolyticus by iTRAQ TIAN Juan, YE Chen, LIU Yu-fei, WANG Li, ZHONG Qing-ping (College of Food Science, South China Agricultural University, Guangzhou 510642, China) Abstract: Vibrio parahaemolyticus was induced into viable but nonculturable (VBNC) state by food preservative at low temperature and oligotrophic condition. The aim of this study is to screen differentially expressed proteins of VBNC state and exponential phase of V. parahaemolyticus using isobaric rags for relative and absolute quantitation (iTRAQ) combined with LC/MS. Functional annotations of unigenes were analyzed using protein sequence similarity, KEGG pathway and GO analysis. The results showed that only 40~45 days V. parahaemolyticus could induce into VBNC state at 4 ℃in seawater containing 10 mmol/L potassium sorbate. A total of 135,880 pr