【作者】 李鸿岩； 赵兴绪； 张红； 王燕玲； 李发弟； 马友记； 张勇；

【Author】 LI Hong-yan~1,ZHAO Xing-xu~2,ZHANG Hong~3,WANG Yan-ling~3, Li Fa-di~4,Ma You-ji~4,ZHANG Yong~(2*) (1 College of Life Sciences and Technology.Gasu Agricultural University,Lanzhou 730070; 2 College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070; 3 Institute of Modern Physics,Chinese Academy of Sciences,Lanzhou 730000; 4 College of Animal Science,Gansu Agriculture University,Lanzhou 730070)

【机构】 甘肃农业大学生命科学技术学院； 甘肃农业大学动物医学院； 中国科学院近代物理研究所； 甘肃农业大学动物科技学院；

【摘要】 (目的)为了获得绵羊精子蛋白质二维电泳图谱,并对0.5Gy和3Gy剂量重离子辐射绵羊精子蛋白质做差异分析.(方法)本研究比较了热的Trizol法制备蛋白质不同上样量对绵羊精子蛋白质二维电泳图谱的影响,利用PDQuest 8.0分析0.5Gy和3Gy剂量2D图谱差异蛋白质点.(结果)结果显示:二维电泳图经硝酸银染色、PDQuest 8.0分析,合适的上样量为0.35mg(pH4～7),蛋白质分子量基本分布在25～66.2kDa.差异凝胶电泳揭示绵羊精子蛋白质经0.5Gy和3Gy剂量重离子辐射后有质和量的改变:其中0.5Gy剂量新表达的蛋白质点有5个,缺失的有1个,表达上调的有3个,表达下调的有2个;3Gy剂量新表达的蛋白质点有8个,缺失的有1个,表达上调的有4个,表达下调的有3个.作为一项阶段性的实验成果,研究表明,热的Trizol法制备精子蛋白结合合适的上样量能够建立绵羊精子全蛋白质图谱,研究发现的重离子辐射引起差异表达蛋白质点为进一步研究重离子辐射绵羊精子蛋白质组学奠定了基础.更多还原

【Abstract】 (Objective)In order to obtain sheep sperm 2-dimensional gel electrophoresis maps,make analysis of 0.5Gy dose and 3Gy dose heavy ion radiation of sheep sperm proteins.(Method) the total protein number of samples processed by hot Trizol and combined different loading quantities on 2-DE map were compared.The different protein spots were analyzeed with PDQuest 8.0 in 0.5Gy dose and 3Gy dose 2D gel. (Result) The 2D gels were stained with silver staining and were analyzsed with PDQuest 8.0 showed that the appropriate loading quantity is 0.35mg(pH4～7),the molecular weight of the most proteins were from 25 to 66.2kDa.The differential in 2 gel of 0.5Gy dose and 3Gy dose showed that 5 protein spots were new expression,1 spots were absent,3 spots increased and 2 spots decreased in 0.5Gy dose gel,8 protein spots were new expression,1 spots were absent,4 spots increased and 3 spots decreased in 3Gy dose gel.The results indicated that many proteins change before and after heavy ion radiation.It was concluded that the protein map of sheep sperm proteome could be constructed with the hot Trizol method and combining with appropriate loading quantities.The differential proteins found after heavy ion radiation of sheep sperm might be used as a plantform for further studies on heavy ion radiation of sheep sperm proteomic.更多还原