5种德国小蠊全虫蛋白提取方法的效果比较  

Comparison of five methods of protein extraction from Blattella germanica

作　　者：黄艳红[1] 霍新北[2,3] 王钰雅[1] 张凡[1] 

机构地区：[1]山东师范大学生命科学学院,山东济南250014 [2]山东省疾病预防控制中心病媒生物防制所 [3]山东省传染病预防控制重点实验室

出　　处：《中国媒介生物学及控制杂志》2012年第3期209-211,共3页Chinese Journal of Vector Biology and Control

基　　金：山东省教育厅科技发展计划项目(J11LC13);山东省医药卫生科技发展计划资助项目(2011HZ056)~~

摘　　要：目的寻找一种适合德国小蠊全虫蛋白提取的方法,为德国小蠊蛋白质组研究提供依据。方法以丙酮为溶剂对德国小蠊雌雄成虫进行脱脂,并利用TCA/丙酮、RIPA裂解液、TrisHCl、1%SDS和裂解液5种蛋白提取液提取全虫蛋白,通过Bradford法测定蛋白浓度,十二烷基磺酸钠-聚丙烯酰胺凝胶电泳比较脱脂前及脱脂后各种蛋白提取方法的提取效果。结果脱脂处理有助于德国小蠊成虫蛋白的提取;5种蛋白提取方法中,RIPA裂解液对2种成虫蛋白提取效果最好,电泳后雌雄成虫蛋白条带分别为13条和16条,其次分别为1%SDS、TrisHCl及裂解液,TCA/丙酮提取法效果最差。结论丙酮脱脂可以明显提高德国小蠊成虫蛋白的提取效果;RIPA裂解液相对更适合于提取德国小蠊雌雄成虫的全虫蛋白质。Objective To explore a suitable method of protein extraction from German cockroach （Blattella germanica） in order to provide a basis for proteomic studies of B. germanica. Methods The adults of B. germanica were defatted with acetone and five protein extracts （TCA/acetone, RIPA, Tris-HCl, 1% SDS and lysate） were used for protein extraction. The concentration of protein was measured with the Bradford methods and the efficacy of protein extraction of the individual method before and after defatting was compared by SDS-PAGE analysis. Results Defatting contributed to the extraction of protein from B. germanica. In the five protein extraction methods, RIPA lysate was most effective. The results showed that the protein bands of female and male adults were 13 and 16, respectively, followed by the methods of SDS, Tris-HCl and lysate, with the TCA/acetone method being the least effective in terms of protein extraction. Conclusion Defatting with acetone makes for protein extraction from B. germanica; and RIPA lysate is more effective for protein extraction from adult B. germanica.

关 键 词：德国小蠊 提取方法 蛋白浓度 十二烷基磺酸钠-聚丙烯酰胺凝胶电泳 

分 类 号：R384.9[医药卫生—医学寄生虫学]