青海湖裸鲤同工酶表达的组织特异性分析

The specific expression of isozymes in different tissues of Qinghai Lake naked carp Gymnocypris przewalskii Przewalskii

使用水平淀粉凝胶电泳方法,分析了20尾青海湖裸鲤的背部肌肉、肝脏、心脏和肾脏4种不同组织的13种同工酶(LDH,MDH,ME,CAT,IDH,EST,ACP,POD,HK,PGM,GDH,SOD,GPI)差异表达,并对部分同工酶基因位点及表达酶谱表型进行了初步分析,以期为其种质资源保护和开发以及遗传育种等方面的研究提供基础资料。结果显示,13种酶类中12种在4种组织中表现出明显的组织差异性,仅有ME在4种组织间差异性较小。其中HK以及GDH的组织差异性尤为明显,仅在肝脏组织中表达。同时使用了适用于这13种酶类表达的3种缓冲系统(TC8.0、TC7.0、EBT),共检测到26个基因位点。其中,Cat,Pgm,Est-1,Gdh,Hk和s-Mdh等6个基因位点为多态位点。进一步分析了这6种多态位点酶类的亚基类型、多态基因位点以及基因位点命名,并计算了多态位点比例(P0.99)为23.08%。

Twenty Qinghai Lake naked carp Gymnocypris przewalskii Przewalskii samples were collected from Qinghai Lake in July 2005. Horizontal starch gel electrophoresis was used to detect the expression of thirteen isozymes （LDH, MDH, ME, GPI, CAT, IDH, EST, ACP, POD, HK, PGM, GDH, and SOD） in four tissues, including muscle, liver, heart and kidney. The results showed that the electrophoresis pattern of twelve isozymes exhibited an apparent tissue-specificity except ME, and the HK, GDH, ACP isozymes showed more remarkable tissue specificity. Three kinds of buffer systems（TC7.0, TCS. 0 and EBT） were tested. Among which, TC7.0 showed good resolution for the three isozymes of HK,PGM and GPI. EBT was superior for EST, and TC8.0 was good for expression of LDH, MDH, ME, ACP, SOD, IDH, POD, GDH and CAT. Biochemical genetic analysis showed that the thirteen isozymes were coded by 26 gene loci, six of which were found to be polymorphic. Finally, based on the zymograms, six isozymes including Cat, Pgm, Est-1, Gdh, Hk, s-Mdh were analyzed genetically for their subunit structure, coding gene loci and numbers as well as their specific expression in the Qinghai Lake naked carp, and the proportions of polymorphic loci （P0.99） was 23.08%.