黏液型与非黏液型铜绿假单胞菌的比较蛋白组学分析

发布时间：2018-10-29 19:27

【摘要】：目的探讨黏液型与非黏液型铜绿假单胞菌(Pseudomonas aeruginosa,P.a)的差异蛋白谱,以发现黏液型P.a潜在的生物标志物。方法采用来自于临床囊性纤维化患者分离的3株黏液型P.a菌株,脉冲场凝胶电泳以确定它们的同源性;采用同位素相对和绝对定量标记(isobaric tags for relative and absolute quantitation,i TRAQ)结合二维液相色谱串联质谱(two-dimensional liquid chromatography-tandem mass spectrometry,2D LC-MS/MS)技术分析3株黏液型与对照组1株非黏液型P.a的差异蛋白谱,并采用RT-q PCR方法对5个共同差异蛋白进行验证。结果 3株黏液型P.a菌株不具有同源性,i TRAQ结果没有明显差异。3组共有219个上调和101个下调差异蛋白。pa2815、pa2018、dnak、pa5046和acef基因转录水平与i TRAQ结果一致。结论不同黏液表型P.a的i TRAQ结果没有明显差异,219个上调和101个下调差异蛋白可以作为研究黏液型与非黏液型P.a致病机制的候选生物标志物。
[Abstract]:Objective to investigate the differential protein profiles of mucinous and non-mucous Pseudomonas aeruginosa (Pseudomonas aeruginosa,P.a) and to identify potential biomarkers of mucinous P.a. Methods three myxotypic P.A strains isolated from patients with clinical cystic fibrosis were used to identify their homology by pulsed field gel electrophoresis. Two dimensional liquid Chromatography tandem Mass Spectrometry (two-dimensional liquid chromatography-tandem mass spectrometry,) with Isotope relative and absolute quantitative labeling (isobaric tags for relative and absolute quantitation,i TRAQ 2D LC-MS/MS) technique was used to analyze the differential protein profiles of three strains of mucus type and one strain of non-mucus type P.A in control group. Five common differentially expressed proteins were verified by RT-q PCR method. Results there was no significant difference in, i TRAQ results among 3 mucinous P.A strains without homology. There were 219 up-regulated and 101 down-regulated differentially expressed proteins in the three groups. The transcription levels of pa2815,pa2018,dnak,pa5046 and acef genes were consistent with those of I TRAQ. Conclusion there is no significant difference in I TRAQ results among different mucotypic phenotypes. 219 up-regulated and 101 down-regulated differentially expressed proteins can be used as candidate biomarkers for studying the pathogenetic mechanism of mucinous and non-mucinous P.A.
【作者单位】：广东医科大学附属医院检验科;深圳市疾病预防控制中心毒理实验室;广东医科大学附属医院临床研究中心;
【基金】：国家自然科学基金青年科学基金(81301478) 广东省湛江市非资助科技计划项目(2015B01042) 广东省医学科学技术研究基金项目(B2017055)~~
【分类号】：R446.5

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