高粒重(48号)和低粒重(154号)小麦株系是01-35(大粒)和6044(小粒、多穗)的稳定杂交后代,具有相似的遗传背景.取灌浆期(开花后17天)的小麦籽粒,提取代谢蛋白,双向电泳后,进行图谱分析.结果显示:两品系中分别鉴定了380和367个蛋白点,其中5...高粒重(48号)和低粒重(154号)小麦株系是01-35(大粒)和6044(小粒、多穗)的稳定杂交后代,具有相似的遗传背景.取灌浆期(开花后17天)的小麦籽粒,提取代谢蛋白,双向电泳后,进行图谱分析.结果显示:两品系中分别鉴定了380和367个蛋白点,其中52个蛋白点为两品系差异显著性表达.将其中13个差异点采用基质辅助激光解析电离飞行时间质谱(matrix assisted laser desorption/ionizaton time of flight mass spectrometry,MALDI-TOF-MS)进行了肽质指纹图分析,通过采用Mascot软件对MSDB数据库查询,其中9个蛋白质点得到了鉴定,包括葡聚糖合成酶、葡萄糖-1-磷酸-腺苷转移酶小亚基前体、一种胚胎晚期富集蛋白等.本研究初步建立了小麦籽粒灌浆其代谢蛋白蛋白质组分析的双向电泳(2-DE)方法,通过质谱分析、数据库搜索的结果对粒重形成的机制进行了探讨,并对以后的研究工作提出了改进方案.展开更多
AIM:To explore the method for early diagnosis of gastric cancer by screening the expression spectrum of saliva protein in gastric cancer patients using mass spectrometry for proteomics.METHODS:Proportional peptide mas...AIM:To explore the method for early diagnosis of gastric cancer by screening the expression spectrum of saliva protein in gastric cancer patients using mass spectrometry for proteomics.METHODS:Proportional peptide mass fingerprints were obtained by analysis based on proteomics matrix-assisted laser desorption ionization time-of-flight/mass spectrometry.A diagnosis model was established using weak cation exchange magnetic beads to test saliva specimens from gastric cancer patients and healthy subjects.RESULTS:Significant differences were observed in the mass to charge ratio(m/z) peaks of four proteins(1472.78 Da,2936.49 Da,6556.81 Da and 7081.17 Da) between gastric cancer patients and healthy subjects.CONCLUSION:The finger print mass spectrum of saliva protein in patients with gastric cancer can be established using gastric cancer proteomics.A diagnostic model for distinguishing protein expression mass spectra of gastric cancer from non-gastric-cancer saliva can be established according to the different expression of proteins 1472.78 Da,2936.49 Da,6556.81 Da and 7081.17 Da.The method for early diagnosis of gastric cancer is of certain value for screening special biological markers.展开更多
Disulfide bond formation protein A （DsbA） is one of the important helper proteins for folding in protein synthesis in vivo. In this study, purification of recombinant DsbA was investigated by examining four importan...Disulfide bond formation protein A （DsbA） is one of the important helper proteins for folding in protein synthesis in vivo. In this study, purification of recombinant DsbA was investigated by examining four important factors with Box-Behnken design method, a statistic-based design of experiments. The optimal operation conditions were obtained by adopting the effectiveness coefficient method on the multi-objective problem, which takes the protein recovery, purification efficiency and throughput of ion-exchange chromatography into account. After the optimization, protein recovery of 96.8% and purity higher than 95% DsbA was achieved, and the productivity was （377.9±1.7） mg soluble DsbA per liter broth. The purified protein was identified by peptide mass fingerprinting matching the record of gil2624856, a mutant of DsbA. The DsbA was preliminarily applied to the refolding of denatured lysozyme in vitro.展开更多
To establish two-dimensional electrophoresis profiles with high resolution and reproducibility from murine brain tissues by human cytomegalovirus（HCMV） infection and paired murine brain tissues and to identify the d...To establish two-dimensional electrophoresis profiles with high resolution and reproducibility from murine brain tissues by human cytomegalovirus（HCMV） infection and paired murine brain tissues and to identify the differential expression proteins. Methods： Forty Kunming mice were randomly divided into infection group （20） injected with HCMVAD169 and control group （20） injected with saline into their brain. After 30 days, the murine brain tissues by HCMV infection and paired murine brain tissues were separated by two-dimensional gel electrophoresis（2-DE）, analyzed by Image Master 2D software, and identified by peptide mass fingerprint（PMF） and database searching, and make Western blotting analyses the differential expression of individual proteins. Results： Well resolved, reproducible 2-D maps of the above tissues were obtained. Some of the different proteins identified by mass spectrometry（MS） were matched in the SWISS-2D PAGE database, Western blotting analyses were further carried out to verify the differential expression of individual proteins. Conclusion： These data will be valuable for studying the diagnosis of disease at an early stage, mechanisms of pathogenic and the key to the development of anti-HCMV medicine.展开更多
文摘高粒重(48号)和低粒重(154号)小麦株系是01-35(大粒)和6044(小粒、多穗)的稳定杂交后代,具有相似的遗传背景.取灌浆期(开花后17天)的小麦籽粒,提取代谢蛋白,双向电泳后,进行图谱分析.结果显示:两品系中分别鉴定了380和367个蛋白点,其中52个蛋白点为两品系差异显著性表达.将其中13个差异点采用基质辅助激光解析电离飞行时间质谱(matrix assisted laser desorption/ionizaton time of flight mass spectrometry,MALDI-TOF-MS)进行了肽质指纹图分析,通过采用Mascot软件对MSDB数据库查询,其中9个蛋白质点得到了鉴定,包括葡聚糖合成酶、葡萄糖-1-磷酸-腺苷转移酶小亚基前体、一种胚胎晚期富集蛋白等.本研究初步建立了小麦籽粒灌浆其代谢蛋白蛋白质组分析的双向电泳(2-DE)方法,通过质谱分析、数据库搜索的结果对粒重形成的机制进行了探讨,并对以后的研究工作提出了改进方案.
基金Supported by The National Natural Science Foundation of China,No. 30640071
文摘AIM:To explore the method for early diagnosis of gastric cancer by screening the expression spectrum of saliva protein in gastric cancer patients using mass spectrometry for proteomics.METHODS:Proportional peptide mass fingerprints were obtained by analysis based on proteomics matrix-assisted laser desorption ionization time-of-flight/mass spectrometry.A diagnosis model was established using weak cation exchange magnetic beads to test saliva specimens from gastric cancer patients and healthy subjects.RESULTS:Significant differences were observed in the mass to charge ratio(m/z) peaks of four proteins(1472.78 Da,2936.49 Da,6556.81 Da and 7081.17 Da) between gastric cancer patients and healthy subjects.CONCLUSION:The finger print mass spectrum of saliva protein in patients with gastric cancer can be established using gastric cancer proteomics.A diagnostic model for distinguishing protein expression mass spectra of gastric cancer from non-gastric-cancer saliva can be established according to the different expression of proteins 1472.78 Da,2936.49 Da,6556.81 Da and 7081.17 Da.The method for early diagnosis of gastric cancer is of certain value for screening special biological markers.
基金Supported by the National Natural Science Foundation of China （21036005）.
文摘Disulfide bond formation protein A （DsbA） is one of the important helper proteins for folding in protein synthesis in vivo. In this study, purification of recombinant DsbA was investigated by examining four important factors with Box-Behnken design method, a statistic-based design of experiments. The optimal operation conditions were obtained by adopting the effectiveness coefficient method on the multi-objective problem, which takes the protein recovery, purification efficiency and throughput of ion-exchange chromatography into account. After the optimization, protein recovery of 96.8% and purity higher than 95% DsbA was achieved, and the productivity was （377.9±1.7） mg soluble DsbA per liter broth. The purified protein was identified by peptide mass fingerprinting matching the record of gil2624856, a mutant of DsbA. The DsbA was preliminarily applied to the refolding of denatured lysozyme in vitro.
基金Supported by the Program of Science and Technology from Shenzhen City, Guangdong Province (200802051)
文摘To establish two-dimensional electrophoresis profiles with high resolution and reproducibility from murine brain tissues by human cytomegalovirus（HCMV） infection and paired murine brain tissues and to identify the differential expression proteins. Methods： Forty Kunming mice were randomly divided into infection group （20） injected with HCMVAD169 and control group （20） injected with saline into their brain. After 30 days, the murine brain tissues by HCMV infection and paired murine brain tissues were separated by two-dimensional gel electrophoresis（2-DE）, analyzed by Image Master 2D software, and identified by peptide mass fingerprint（PMF） and database searching, and make Western blotting analyses the differential expression of individual proteins. Results： Well resolved, reproducible 2-D maps of the above tissues were obtained. Some of the different proteins identified by mass spectrometry（MS） were matched in the SWISS-2D PAGE database, Western blotting analyses were further carried out to verify the differential expression of individual proteins. Conclusion： These data will be valuable for studying the diagnosis of disease at an early stage, mechanisms of pathogenic and the key to the development of anti-HCMV medicine.
