杜氏盐藻高盐诱导蛋白的分离和鉴定

摘要

目的:对高盐和低盐培养条件下杜氏盐藻蛋白质组的双向电泳(2-DE)图谱进行比较分析,筛选高盐诱导蛋白并鉴定.方法:提取高盐(3.5 mol/L NaCl)和低盐(1.5 mol/L NaCl)培养条件下杜氏盐藻的总蛋白并进行2-DE,考马斯亮蓝染色后用Image-Scanner扫描仪扫描,ImageMaster 5.0图像分析软件分析,并利用基质辅助激光解析电离质谱法(MALDI-TOF)对在高盐培养条件下增强表达达到3倍的蛋白进行鉴定.结果:2-DE图谱上共分离出509个杜氏盐藻蛋白质斑点,主要分布在等电点4～8和相对分子质量20000～97000的范围内;高盐培养条件下有21个蛋白增强表达达到3倍并且通过MALDI-TOF鉴定出了其中的12个.结论:成功获得了分辨率和重复性较好的杜氏盐藻蛋白质组2-DE图谱,鉴定得到了12种高盐诱导增强表达达到3倍的蛋白.%Aim: To analyze the two-dimensional electrophoresis(2-DE) maps of Dunaliella salina(D. Salina) under different Na + concentration (3.5 mol/L NaCl and 1.5 mol/L NaCl) and screen differently expressed proteins between them. Methods :2-DE was taken to separate the total proteins from D. Salina treated with high and low concentration salt. Protein patterns were analyzed and identified by ImageMaster 5. 0 software and MALDI-TOF after staining and scanning. Results; A total of 509 protein spots were found as differently expressed proteins,out of which, the expression level of 21 proteins cultured in high concentration salt condition was three times of those cultured in low concentration salt condition.Conclusion: 2-DE profiles from D. Salina with high resolution and reproducibility have been obtained and 12 up-regulated proteins in high salt concentration have been identified.