作　　者： ; ; ; ; ; ; ; ;

机构地区： 华南农业大学动物科学学院蚕丝科学系

出　　处： 《中国农业科学》 2008年第11期3933-3940,共8页

摘　　要： 【目的】用盐酸刺激活化家蚕滞育卵的孵化,是蚕种生产长期沿用的一项人工孵化技术,但盐酸能活化滞育卵的机制却有许多未明之处,为此,从蛋白质组学的角度探讨盐酸对蚕卵滞育解除的作用关系。【方法】用双向电泳技术和电喷雾串联质谱技术,比较分析冷藏45d的滞育卵浸酸前后的难溶性卵蛋白的差异表达。【结果】根据双向电泳图谱分析,在浸酸前的图谱中共检测到296个蛋白点,浸酸后的检测到302个蛋白点,两者相匹配的蛋白斑点有265个,匹配率为88.6%。特异性的蛋白点,浸酸前有31个,浸酸后有37个。对浸酸前存在、经浸酸处理后消失的2个特异性差异的高丰度蛋白点用电喷雾串联质谱技术作分析,NO.1蛋白序列与浆膜蛋白(chorion protein[Bombyx mori])有55个氨基酸的肽段相匹配,NO.2蛋白序列与热激蛋白(heat shock protein hsp19.9[Bombyx mori])只有15个氨基酸的肽段相匹配,推测是一种未知蛋白。【结论】滞育性蚕卵经浸酸后,难溶性卵蛋白有差异表达,浸酸前后的一些蛋白种类或分子量发生了变化。 [Objective] Using HC1 to activate the diapause eggs is a traditional technique of artificial hatching applied in silkworm egg production. Its mechanism has not yet been clarified. This experiment explored the effect of HCI on the termination of diapause of silkworm eggs cold-stored for 45 days from the point of proteomes. [Method] Two-dimensional electrophoresis techniques and ESI-MS-MS were used to compare and analyze the variation expression of difficult-dissolved-proteins of diapause eggs cold-stored for 45 days before and after acid treatment. [Result] Through analysis on the two-dimensional electrophoretogram, there were 296 and 302 dots were found before and after acid treatment, respectively. Among them 265 dots were matchable. The matchability reached 88.6%. There were 31 and 37 specific protein dots were found before and after acid treatment, respectively. ESI-MS-MS analysis was conducted for 2 specific protein-rich dots which disappeared after acid treatment. The results showed that the sequence of No.1 protein dot had 55 amino acid peptides matched with those of chorion protein [Bombyx rnori]. While the sequence of No.2 protein dot had only 15 amino acid pepides matched with those of heat shock protein （hsp 19.9 [Bombyx mori]）, and it was presumed to be an unknown protein. [Conclusion] The difficult-dissolved-proteins of diapause eggs have variation expression after acid treatment. Some proteins before and after acid treatment are changed in MW.

关 键 词： 家蚕卵 浸酸 蛋白质 电泳 图谱分析

领　　域： [农业科学] [农业科学] [生物学]