CAJ | 학술논문

目的利用双向电泳和质谱鉴定技术探讨环境和人体中霍乱弧菌蛋白表达的差异.方法利用适当的裂解液处理霍乱弧菌,提取全菌蛋白;采用pH梯度等电聚焦对全菌蛋白进行双向电泳;考马斯亮蓝染色后获得双向电泳图谱,并利用ImageMaster 2D Elite 5.0图像分析软件进行分析,所得的数据采用SPSS15.0进行统计分析,找出差异蛋白;在此基础上,胰蛋白酶消化这些特殊差异蛋白,并进行质谱(MALDI-TOF-MS)分析.结果获得1032±22个蛋白斑点,蛋白主要集中在等电点(PI)4.00～7.20之间,重复胶的匹配点数为1025±24,匹配率为96.30%;发现了有明显差异的21个蛋白点,并用质谱鉴定了其中4种蛋白.结论获得了环境和人体中霍乱弧菌蛋白的差异表达蛋白.
목적 이용쌍향전영화질보감정기술탐토배경화인체중곽란호균단백표체적차이.방법 이용괄당적렬해액처리곽란호균,제취전균단백;채용pH제도등전취초대전균단백진행쌍향전영;고마사량람염색후획득쌍향전영도보,병이용ImageMaster 2D Elite 5.0도상분석연건진행분석,소득적수거채용SPSS15.0진행통계분석,조출차이단백;재차기출상,이단백매소화저사특수차이단백,병진행질보(MALDI-TOF-MS)분석.결과 획득1032±22개단백반점,단백주요집중재등전점(PI)4.00～7.20지간,중복효적필배점수위1025±24,필배솔위96.30%;발현료유명현차이적21개단백점,병용질보감정료기중4충단백.결론 획득료배경화인체중곽란호균단백적차이표체단백.
Objective To study the different proteins of human and environment Vibrio cholerae (V. cholerae) by two dimensional electrophoresis (2-DE) and TOF-TOF-MS. Methods Total cellular pro-tein was extracted by lysate from V. cholerae, and proteins were separated by 2-DE under immobilized pH gradients(IPG), then electrophoregrams were dealed with coomassie brilliant blue, and analyzed by Im-ageMaster 2D Elite 5.0, finally the different protein spots were identified by TOF-TOF-MS. Results High repetitive 2-DE maps were obtained. 2-DE and image analysis revealed 1032±22 protein spots, and PI value was among 4.00 to 7.20. Matching spots were 1025±24 in two repeats electrophoregrams and matching ratio was 96.30%. Conclusion The different protein spots were successfully established with high quality and sharpness separation by 2-DE and TOF-TOF-MS, which stands as a valuable resource for proteomics re-search of V. cholerae.