Isozymes Analysis of the Golden Cuttlefish Sepia esculenta（Cephalopoda ： Sepiidae）

Thirty-nine isozymes in four tissues (mantle muscle, buccal bulb muscle, eye and liver) of Sepia esculenta were screened for enzymatic analysis using starch gel electrophoretic technique. Eighteen enzymes (G3PDH, LDH, MDH, MEP, IDHP, PGDH, GRS, NP, AAT, CK, AK, EST, ALP, ACP, FBP, MPI, GPI and PGM) show strong activities and good convergence in zymogram. They are proved to be suitable genetic markers in Sepia esculenta. Among the tissues used, mantle muscle is the best for electrophoretic analysis of isozymes. Eye and liver are fairly good for some special enzymes, such as LDH, EST, MPI, etc. Twenty-six loci are detected. The proportion of polymorphic loci is 0.115 in the Qingdao sample and 0.153 in the Rizhao sample (P<0.99). The mean values of the observed and expected heterozygosity per locus of Qingdao sample are 0.016 and 0.017, while those of the Rizhao sample are 0.023 and 0.025 respectively.     

Antioxidant and isozyme features of two strains of Laminaria japonica （Phaeophyceae）

Healthy sporophytes of two gametophyte mutants of Laminaria japonica with different heat resistances: kelp 901 (901, with comparatively stronger heat-resistance) and Rongcheng No.1 (RC, sensi- tive to heat stress), were respectively collected during October to December 2002 from Yantai and Rong- cheng Sea Farm in the Shandong Peninsula of China. The contents of some biochemical materials and antioxidant capacity were analyzed under controlled laboratory conditions to identify if there is any rela- tion between the overall antioxidant capacity and the heat-resistance in L. japonica and to understand possible mechanism of heat-resistance. Results show that: (1) the overall antioxidant capacity in healthy sporophyte of 901, such as vitamin E, polyphenol, and ascorbic acid contents and the enzymatic activity of SOD, POD, CAT, Gpx, PPO, and PAL, were not always higher than that of RC under controlled labo- ratory conditions, and no significance (P>0.05) was shown in total antioxidant capacity (T-AOC) in 901 and RC. Result suggested that the difference in antioxidant capacity was not a decisive factor for different heat-resistances in L. japonica; (2) the simultaneous assay on isozymes was carried out using vertical polyacrylamide gel electrophoresis (PAGE). Considerable differences in peroxide (PRX), malate dehy- drogenase (MDH), malic enzyme (ME), polyphenol oxidase (PPO) and glutamate dehydrogenase (GDH) were obtained in 901 and RC from either the band number, relative mobility (Rf), or staining intensity, and ME could be used as an indicator to distinguish healthy sporophyte of 901 and RC under controlled labo- ratory conditions.     

管角螺不同组织同工酶的研究

采用聚丙烯酰胺垂直板梯度凝胶电泳技术，对管角螺7种组织（外套膜、肌肉、肝脏、心脏、生殖腺和缠卵腺）的5种同工酶（EST、SOD、ME、MDH、LDH）进行了分析，结果表明：5种同工酶存在不同程度的组织特异性，SOD、EST、LDH在肝脏组织中有较高活性，而MDH、ME在心脏组织中活性较高。     

似刺鳊鮈发育过程中同工酶变化及组织分布特征分析

运用高pH不连续聚丙烯酰胺凝胶(PAGE)电泳技术分析了似刺鳊鮈胚胎、仔鱼发育过程中5种同工酶变化,并对成鱼的9种组织同工酶分布特征进行了检测。结果发现:除醇脱氢酶(ADH)外,其他4种酶(EST、LDH、MDH、SOD)在发育过程中均被检出,且呈现明显的阶段性特征;在成鱼的4种同工酶也存在明显的组织特异性。4种酶共检测到9个基因位点。     

中华绒螯蟹不同组织LDH同工酶凝胶电泳分析

采用高pH不连续系统聚丙烯酰胺垂直板凝胶电泳,特异性组织化学染色对中华绒螯蟹(Eriocheir sinensis)的血清、肌肉、鳃、心脏、肝胰腺、精巢和精子等组织的乳酸脱氢酶(LDH)同工酶酶谱进行了研究。同时,利用2种底物(乳酸钠和α-羟基丁酸钠)进行了特异性反应,并对其精子特异性乳酸脱氢酶进行了研究。结果表明:以乳酸钠为底物时,中华绒螯蟹的肌肉、鳃、血清和心脏有2条谱带,而精巢和精子有3条谱带,但在肝胰腺中未发现谱带;以α-羟基丁酸钠为底物时,只在血清、精巢和精子发现有1条谱带。从而说明,中华绒螯蟹乳酸脱氢酶同工酶存在组织特异性;精子存在特异性乳酸脱氢酶。     

月鳢个体发育早期4种同工酶的初步研究

采用垂直板聚丙烯酰胺凝胶电泳技术,对月鳢早期(从未受精卵至卵黄吸尽期)8个发育阶段的LDH、MDH、ADH、EST同工酶进行研究。结果表明:月鳢的早期发育阶段ADH酶谱相对稳定,均有两条谱带;LDH、MDH和EST三种同工酶的酶谱在神经胚期前均无明显变化,此后随发育进程逐渐复杂。     

大弹涂鱼不同组织器官的同工酶研究

采用垂直聚丙烯酰胺平板电泳技术,对大弹涂鱼(Boleophthalmus pectinirostris)的眼睛、心脏、肝脏、肌肉、脾脏和鳃等6种组织器官中的13种同工酶进行研究,并分析了其酶谱表型。结果表明,四唑氧化酶(TO)和酸性磷酸酶(ACP)没有检测到酶带;三梨醇脱氢酶(SDH)和葡萄糖-6-磷酸脱氢酶(G-6-PDH)仅在肝脏中表达;超氧化物歧化酶(SOD)在所有组织中均存在,部分组织中存在差异;乳酸脱氢酶(LDH)、苹果酸脱氢酶(MDH)、苹果酸酶(MEP)、酯酶(EST)、醇脱氢酶(ADH)、D-葡萄糖脱氢酶(GcDH)、甲酸脱氢酶(FDH)、谷氨酸脱氢酶(GDH)则具有明显的组织特异性。     

三疣梭子蟹肌肉乳化病的同工酶病理变化研究

采用垂直平板聚丙烯酰胺凝胶电泳技术(PAGE),对患有肌肉乳化病的三疣梭子蟹(Portunus trituberculatus)3种组织中10种同工酶及肌浆蛋白的酶谱表型进行分析,并就患病后酶谱变化特征及原因进行了讨论。证实在部分组织中,除SOD酶外9种酶及肌浆蛋白的酶带均出现明显的紊乱现象,表明病蟹的病理机制极可能与代谢异常有关,其特异性变化可以作为疾病早期诊断及病理分析的生化辅助指标。     

尖刀蛏和小荚蛏不同组织同工酶的比较分析

通过聚丙烯酰胺凝胶垂直板不连续电泳对尖刀蛏外套膜、鳃、肝胰腺和足等4种组织和小荚蛏外套膜、足和内脏团等3种组织的9种同工酶进行研究。结果表明,两种蛏相应组织不同同工酶的分布既具一定组织特异性,也存在一定种属特异性,且两种蛏的种属差异明显大于同种蛏的组织间差异。其中,尖刀蛏与小荚蛏在足组织间差异最大,而尖刀蛏肝胰腺与小荚蛏内脏团两组织间的差异相对较小。由于不同种蛏各具其稳定的酶谱表型,故可利用两种蛏同工酶酶谱表型的特异性作为蛋白质分子标记而应用于刀蛏科不同属蛏的物种鉴定。     

不同年龄泥蚶同工酶电泳的研究

采用聚丙烯酰胺凝胶电泳方法，对同一种群不同年龄的泥蚶过氧化物酶、酯酶、淀粉酶、苹果酸脱氢酶、过氧化氢酶、醇脱氢酶、苹果酸酶和超氧化物歧化酶等8种同工酶进行了分析比较。结果表明，不同年龄泥蚶的过氧化物酶、酯酶同工酶在酶谱带上有一定差异。淀粉酶、苹果酸脱氢酶、过氧化氢酶、醇脱氢酶、苹果酸酶和超氧化物歧化酶同工酶在酶活性上有一定的差异。