尖尾藻(Oxyrrhis marina)乳酸脱氢酶同工酶的研究  

STUDIES ON LACTATE DEHYDROGENASE ISOENZYMES OF THE MARINE DINOFLAGELLATE(OXYRRHIS MARINA):AN ANALYSIS BY AGAROSE GEL ELECTROPHORESIS

作　　者：郑子修 钟金颜 丁琳华[2] Zheng Zixiu;Zhong Jinyan;Ding Linhua(Kunming Institute of Zoology,Academia Sinica)

机构地区：[1]中国科学院昆明动物研究所 [2]南京生物化学制药研究所

出　　处：《Zoological Research》1988年第S01期33-42,共10页动物学研究（英文）

摘　　要：对培养2至7天分六阶段抽样的尖尾藻细胞(Oxyrrhis marina)进行乳酸脱氢酶同工酶琼脂糖凝胶电泳分析,结果表明,全部样品中乳酸脱氢酶同工酶均呈现三种不同分子形式的表型特征。随电泳迁移率的增大,由阴极趋于阳极端依次为LDH_(3),LDH_(2),LDH_(1),采用分光光度比色定量法,于波长560nm测得各同工酶组份相对百分含量见表1。LDH_(3)同工酶在培养2至7天的全过程中酶活性水平始终保持高于LDH_(1)和LDH_(2)的水平。在培养第四天山现最高值。The glycolytic enzyme lactate dehydrogenase(LDH,EC 1.1.1.27),which catalyzes the reversible conversion of pyruvate to lactate and occurs in virtually all tissues,exists in multiple molecular forms called isoenzymes.Lactate dehydr-ogenase isoenzymes has been extensively studied in invertebrates and vertebrates.The dinoflagellate are a group lower eukaryotic algae possessing a number of unique cellular properties.It is one of the most suitable biological material for the study of origin and evolution from prokaryotcs to eukaryotes.In the present study,the changes of lactate dehydrogenase isoenzymes during different periods of cultured 2 to 7 days in the dinoflagellatc,Oxyrrhis marina were analysed by agarose gel electrophoresis.The algae cells were collected by centri-fugation at 4000 rpm for 15 min and then lyscd with the lysing solution containing 0.3M sucrose,0.025M trihydroxymethylamino,0.04M NaHSO3,0.005M ethylenediamine tetraactic acid,0.01M MgS04 and 0.1%Triton X-100,pH 7.0-7.4.The lysate was centrifuged at 10000 rpm(0°-8℃)for 15 min remove cell debris and nuclei and then supernatant was electrophoresis performed on 0.5%agarose gel thin layer in a pH 8.6 buffer containing 0.06M sodium diethylbarbiturate and 0.2N hydrochloric acid.The LDH isoen-zymic activity in agarose gel was visualized with a staining solution consisting of sodium DL-lactate,Nicotinamide adenine dinucleotide(NAD),N-methylph-enazinium methylsulphate(PMS)and nitro-tetrazolium blue chloride(NBT).The gel was incubated at 37°-40℃for 40 min.The isoenzymic bands were excised from agarose gel and eluted.The electropherograms of lactate dehydrogenase isoenzymes was scanned record by SHIMADZU dual-wavelength chroma-togram scanner model CS-910 atλs=570 nm,λr=500nm and scanning speed for 20mm/min.The results were as follows.(1)The isocnzymic phenotypes of three different molecular forms was observed in 2 to 7 days cultures of Oxyrrhis marina.Different bands are referred to as LDH1,LDH2 and LDH3 in order of relative mobility towards the anode a

关 键 词：涡鞭毛虫 尖尾藻 乳酸脱氢酶同工酶 琼脂糖凝胶电泳 

分 类 号：Q55[生物学—生物化学]