|
|||||
|
|
| “鸟枪”蛋白组学技术鉴定牙龈卟啉单胞菌47A-1亚型外膜蛋白 | |
| 引用本文： | 孙俊毅,孙媛媛,苟建重,李昂.“鸟枪”蛋白组学技术鉴定牙龈卟啉单胞菌47A-1亚型外膜蛋白[J].西安交通大学学报(医学版),2011,32(2):170-174. |
| 作者姓名： | 孙俊毅 孙媛媛 苟建重 李昂 |
| 作者单位： | 1. 西安交通大学医学院附属口腔医院牙周科,陕西西安,710004 2. 烟台市口腔医院牙周科,山东烟台,264001 3. 西安交通大学医学院附属口腔医院口腔医学研究中心,陕西西安,710004 |
| 基金项目： | 国家自然科学基金资助项目(No.30500560)~~ |
| 摘 要： | 目的利用shotgun(鸟枪)蛋白质组学策略鉴定牙龈卟啉单胞菌(Porphyromonas gingivalis,P.g)亚型47A-1外膜蛋白,深化P.g外膜蛋白的蛋白质组学研究,为进一步研究P.g外膜蛋白的致病性奠定基础。方法提取P.g47A-1外膜蛋白,SDS-PAGE电泳分离蛋白,胶内酶解为多肽混合物,应用毛细管高效液相色谱加串联质谱分析,SEQUEST程序进行数据库搜索并鉴定蛋白。结果用考马斯亮蓝法染色凝胶分离的蛋白,发现P.g47A-1蛋白条带主要在14.3~97 ku之间。酶解的肽段经毛细管高效液相色谱分离,得到肽段水平的总离子流图,分析肽段序列后发现P.g47A-1蛋白有623个唯一肽段,鉴定出136种蛋白,包括了51种已知蛋白,85种未知蛋白。结论 Shotgun技术可分析出P.g47A-1大量未知蛋白,尤其是丰度低、疏水性强的外膜蛋白,优于二维凝胶电泳(two dimensional gelelectrophoresis,2-DE)等传统蛋白质组学方法,为进一步研究P.g外膜蛋白的致病性奠定了基础。 |
| 关 键 词： | 牙龈卟啉单胞菌 外膜蛋白 鸟枪法 蛋白质组学 |
Identification of outer membrane proteins of Porphyromonas gingivalis 47A-1 by Shotgun proteomics method |
|
| SUN Jun-yi,SUN Yuan-yuan,GOU Jian-chong,LI Ang.Identification of outer membrane proteins of Porphyromonas gingivalis 47A-1 by Shotgun proteomics method[J].Journal of Xi‘an Jiaotong University:Medical Sciences,2011,32(2):170-174. | |
| Authors: | SUN Jun-yi SUN Yuan-yuan GOU Jian-chong LI Ang |
| Institution: | SUN Jun-yi1,SUN Yuan-yuan2,GOU Jian-chong1,LI Ang3(1.Department of Periodontology,Stomatological Hospital,Medical School ofXi'an Jiaotong University,Xi'an 710004,2.Department of of Periodontolgy,Yantai Stomatological Hospital,Yantai 264001,3.Research Center of Stomatology,Medical School of Xi'an Jiaotong University,Xi'an,710004,China) |
| Abstract: | Objective To identify outer membrane proteins of Porphyromonas gingivalis(P.g.) by shotgun proteomics method so as to lay a foundation for further study of pathogenicity and proteomics of P.g.Methods P.g47A-1 were cultured and collected when they were in good conditions.Outer membrane proteins of Pg 47A-1 were extracted by differential centrifugation and separated by SDS-PAGE.We selected trypsin for protein hydrolysis in SDS-PAGE gel,separated the peptide fragments using cap HPLC,and analyzed each peptide f... |
| Keywords: | Porphyromonas gingivalis outer membrane protein shotgun method proteomics |
| 本文献已被 CNKI 万方数据 等数据库收录！ | |