›› 2000, Vol. 43 ›› Issue (-1): 77-84.

• 研究论文 • 上一篇    下一篇

越冬松针瘿蚊幼虫整体携脂蛋白 的分离和纯化

李毅平1,龚和1,朴镐用2   

  • 出版日期:2000-12-20 发布日期:2000-12-20

Purification and characterization of lipophorin from overwintering mature larvae of pine needle gall midge,Thecodiplosis japonensis (Diptera:Cecidomiidae)

LI Yi-ping, GONG He, Park Ho-yong   

  • Online:2000-12-20 Published:2000-12-20

摘要: 采用KBr密度梯度超速离心并结合常规Sepharose CL-4B凝胶柱层析,从越冬松针瘿蚊Thecodiplosis japonensis(Uchida et Inouye) 幼虫整体中,分离并纯化了一种携脂蛋白。这是第二例从昆虫整体分离并纯化出携脂蛋白的报道。采用凝胶柱层析确定该携脂蛋白的相对分子质量为638 kD,它是由分别为240 kD和52 kD的两个亚基组成 。整体分子中含有52.8%的蛋白和47.2%的脂类 。苏丹黑B和希夫氏试剂染色显示阳性,说明它是一种糖脂复合蛋白。采用超速离心确定它的密度为1.11 g/mL,表明它是一种高密度的脂蛋白。

关键词: 日本松针瘿蚊, 携脂蛋白, 高密度脂蛋白, 越冬幼虫

Abstract: A lipophorin was isolated and purified from whole body of third instar mature larvae of pine needle gall midge,Thecodiplosis japonensis(Uchida et Inouye) by KBr density gradient ultracentrifugation and Sepharose CL-4B gel permeation. It is the second lipophorin that have been purified from whole body. The native molecular weight was 638 kD by gel permeation chromatography. It was composed of two subunits, named as apoLp-Ⅰ and apoLp-Ⅱ, with molecular weight of 240 kD and 52 kD respectively. The holoprotein contained 52.8% protein and 47.2% lipid by weight. Sudan Black B and Periodic acid/Schiffs' reagent staining showed positive results, indicating a kind of glycolipoprotein. Its density was 11.1 g/mL, showing a kind of high density lipoprotein. The presence of high content of lipophorin in wholebody of third instar, suggesting a potential role in lipid transport and/or cryoprotection during the overwintering periods.

Key words: Thecodiplosis japonensis, lipophorin, high density lipoprotein, overwintering larvae