2型猪链球菌分泌组双向电泳样品制备方法的建立  

Improvement of preparation method of Streptococcus suis type 2 secretome sample for two dimensional electrophoresis

作　　者：张爱玲[1] 王兴龙 王英超[1] 党源[1] 张复贤[1] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]军事兽医研究所,吉林长春130062

出　　处：《中国预防兽医学报》2009年第10期776-779,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：973基金资助项目(2006CB504400)

摘　　要：为改进适用于2型猪链球菌分泌组双向电泳样品的制备方法,本研究将无蛋白细菌培养液培养2型猪链球菌,分别以超滤-冻干法、超滤-冻干-丙酮沉淀法和改良超滤-冻干-三氯乙酸(TCA)/丙酮沉淀法富集培养物上清中的细菌分泌蛋白质,通过比较双向电泳的结果优选最佳的样品制备方法。结果证明改良浓缩-冻干-TCA/丙酮沉淀法较为理想,蛋白丢失少、溶解性佳,所得双向电泳图谱清晰,分辨率高。因此,改良浓缩-冻干-TCA/丙酮沉淀法更适用于制备体外培养的2型猪链球菌分泌组双向电泳样品,并可以满足进一步研究的需要。The objective of this experiment is to improve the preparation method of Streptococcus suis type 2 secretome sample for two-dimensional electrophoresis （2-DE）. Streptococcus suis type 2 were cultured in non-protein medium and the protein sample prepared for 2-DE by ultrafiltration-lyophilization, ultrafiltration-lyophilization acetone precipitation and the modified ultrafiltra- tion-lyophilization acid （TCA）/acetone precipitation methods, respectively. The optimized method was determined by comparing 2-DE generated from different preparations. The results indicated that the modified ultrafiltration-lyophilization-TCA/acetone pre- cipitation method produced clear 2-DE images with highest resolution, therefore is the method of choice for preparation of Strepto- coccus suis type 2 secretome 2-DE sample.

关 键 词：2型猪链球菌 分泌组 双向电泳 丙酮沉淀法 TCA/丙酮沉淀法 

分 类 号：S855.1[农业科学—临床兽医学]