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作 者:段志贵[1] 王美迟[1] 胡朝暾[1] 刘珍[1]
机构地区:[1]湖南师范大学生命科学学院,蛋白质化学及发育生物学教育部重点实验室,中国湖南长沙410081
出 处:《生命科学研究》2013年第2期106-110,142,共6页Life Science Research
基 金:国家自然科学青年基金资助项目(31000477);高等学校博士学科点专项科研基金资助课题(20104306120005);湖南省教育厅基金资助项目(10C0961;09C756);湖南师范大学蛋白质化学与发育生物学教育部重点实验室开放基金课题(DF1111;DF1104);湖南师范大学青年科学基金项目(110634)
摘 要:自然界虎纹捕鸟蛛雌蛛数量远多于雄蛛数量,为了探究雌、雄蛛粗毒的差异,用不同方法比较了单个虎纹捕鸟蛛雌、雄蛛粗毒的特征.雌蛛单次螫毒量为(26.5±2.47)μL,冻干后粗毒质量为(5.01±0.78)mg,明显高于雄蛛单次螫毒量(10.83±1.35)μL,冻干后粗毒质量(2.05±0.17)mg;用Lowry法和Bradford法测定雌蛛和雄蛛粗毒的蛋白质含量,两种方法均表明雄蛛粗毒中蛋白含量高于雌蛛.用反相高效液相色谱分离雌蛛和雄蛛粗毒蛋白,并215 nm检测色谱图,发现大部分洗脱峰重叠,而雄蛛色谱图中多两个主峰.Tricine SDS-PAGE电泳分析表明雌蛛粗毒相对分子质量小于10 kD的蛋白质含量较高;而Tris SDS-PAGE电泳分析表明雌蛛粗毒相对分子质量大于10 kD的蛋白质含量较低,基于雌蛛和雄蛛粗毒蛋白含量的差异,有必要对虎纹捕鸟蛛雌、雄蛛粗毒进行独立研究.The number of female spiders Selenocosmia huwena is greater than that of male spiders in nature. In order to investigate the differences in the venom of female and male spiders, the basic characteristics of their venoms were studied by using various methods. Female single sting venom was (26.5±2.47) μL and it was significantly higher than that (10.83±1.35 μL) of male; Lyophilyzed crude venom quantity of a fe- male or male spider was (5.01±0.78) mg and (2.05±0.17) mg, respectively. Using Lowry and Bradford methods to determine the protein content, it was shown that protein content in the venom of male spider was greater than that in the venom of female spider. The female and male spider venoms were separated by using reversed phase high performance liquid chromatography (RP-HPLC) at 215 nm. The elution profiles showed that most of the peaks were overlapped and two unique peaks were present in male's chromatogram. Tricine SDS-PAGE analysis showed female spider venom contained higher content of proteins/peptides with molecular weight less than 10 kD than male spider venom. In contrast, female spider venom was proved by Tris SDS-PAGE analysis to have lower content of proteins with molecular weight greater than 10 kD than male spider venom. Because of the differences, the crude venoms of male and female should be studied respectively.
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