摘要: |
于1994年7—12月,在山东、福建沿海采集真鲷、黑鲷、牙鲆、鲈和小黄鱼等5种鱼63尾692个生化样品,应用水平淀粉胶和垂直聚丙烯酰胺凝胶电泳方法,对5种鱼的肌肉、眼睛、肝脏、心脏、鳍、鳃、肾脏共7种组织的23种同工酶进行初步分析。结果为:1.选出酶活性强,基因座位表达稳定、清晰的肌肉、眼睛、肝脏、心脏4种组织,用于以上5种鱼的常规性生化遗传分析;2.筛选出适于电泳分析的缓冲液系统有TCl,EBT,TG等3种以及14种图谱清晰、分辨率高的同工酶(LDH,MDH,IDHP,MEP,ADH,SDH,G6PDH,CAT,GDH,
PGM,PGDH,GP,EST,Pm,SOD)和肌浆蛋白(Pm);3.对其中的LDH,IDHP,ADH,GDH和BST5种同工酶进行生化遗传分析,包括每种酶的亚基结构、编码座位以及多态座位等位基因数等。 |
关键词: 同工酶 电泳 组织特性 海水鱼 |
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基金项目:国家科委攀登计划B项目,PDB6-5-2 |
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THE SPECIFIC EXPRESSION OF ISOZYMES IN DIFFERENT TISSUES AND PRELIMINARY BIOCHEMICAL GENETIC ANALYSIS BASED ON THE ELECTROPHORESIS OF FIVE MARINE FISHES |
Wang Keling, You Feng, Xu Cheng, Wu Suqi
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Institute of Oceanology, Chinese Academy of sciences, Qingdao 266071
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Abstract: |
Sixty-three fishes belonging to 5 different species (Pagrusonms major, Spams macrocephalus, Paralichthys oliuaceus, Lateolabrax japonicus, Pseudosciaena polyactis) were collected from the coastal waters of Shandong and Fujian provinces from July to December in 1994. Biochemical samples were extracted by homogenizing from seven tissues (muscle, eye, liver, heart, gill, fm and kidney) of each species separately. Horizontal starch gel and vertical polyacrylamide gel electrophoresis were used in examining 692 samples. Four kinds of buffer systems, TC1, TC2 and EBT for starch gel and TG for polyacrylamide gel electrophoresis, were tested. Altogether 22 isozymes and a protein were examined through different kinds of tissue or different kinds of buffer systems. Only EST, Pm, and SOD were electrophoresed by polyacryamide gel, the others by starch gel. The results showed the isozymes expression was highly tissue specific. Among the buffer systems tested, TC1, EBT and TG, seemed very useful for the future application due to their good resolution of isozymes and a protein, which exhibited stable expression and high level of activity in four kinds of tissue (muscle, eye, liver and heart) during the electrophoresis (Tab. 1). Furthermore, two buffer systems supporting the starch gel electrophoresis gave good resolution of 12 isozymes: TCl to LDH, MDH, IDHP and MEP: EBT to ADH, SDH, G6PDH, CAT, GDH, PGM, PGDH, and GPI. Another buffer system, TG was a superior to EST, Pm and SOD for polyacrylarnide gel electrophoresis (Tab.2, Tab.3). Finally, based on the zymograms, five isozymes, e.g. LDH, IDHP, ADH, GDH and EST (Plate I), were analyzed genetically on their subunit structure, coding gene loci and numbers as well as their specific expression in these five marine fishes. |
Key words: Isozyme, Electrophoresis, Tissue specificity, Marine fishes |