双团棘胸蛙与棘腹蛙酯酶同工酶的比较研究

采用垂直聚丙烯酰胺凝胶电泳技术,对六盘水境内的双团棘胸蛙Paa yunnanensis与棘腹蛙P.boulengeri的肝、肠、胃、肺等4种组织中的酯酶(EST)同工酶进行了研究.结果 显示:两种蛙酯酶同工酶共有10条带,其中双团棘胸蛙EST同工酶有9条带,棘腹蛙EST同工酶有7条带,两种蛙EST同工酶存在较大的种间差异,同种蛙不同组织的酯酶(EST)同工酶迁移率、表达强度也不同.双团棘胸蛙酯酶同工酶酶谱较棘腹蛙的复杂.     

意蜂工蜂、雄蜂和蜂王的LDH和EST同工酶比较

用聚丙烯酰胺凝胶电泳的方法,对意蜂工蜂、雄蜂和蜂王以及两种不同大小幼虫之间的乳酸脱氢酶(LDH)和酯酶(EST)同工酶分别作了比较。结果表明,工蜂、雄蜂和蜂王之间LDH同工酶数目无差异,但活性上存在差异;而酯酶同工酶在幼虫阶段与成虫阶段及工蜂、雄蜂和蜂王之间则有着不同的同工酶谱型。说明意蜂同工酶在种内的表达受到发育程度、进食质量、分化方向等因素的影响。     

意蜂和中蜂四种同工酶的研究

用聚丙烯酰胺凝胶等电聚焦电泳分析了意蜂和中蜂的酯酶(Est)、异柠檬酸脱氢酶(Idh)、苹果酸酶(Me)和苹果酸脱氢酶(Mdh)同工酶.两个蜂种的四种同工酶谱有不同程度的差别、意蜂酯酶Ⅳ和苹果酸脱氢酶Ⅲ是多态性的;中蜂的四种同工酶没有多态现象.     

三种微蛛酯酶同工酶的比较

本文对农田常见三种微蛛的酯酶同工酶进行了比较研究,结果表明:三种微蛛的酯酶同工酶共有18条Rf值不同的酶带,但同种不同发育阶段、不同性别的酯酶同工酶带数不同,而对种间来说其酯酶同工酶谱具有种的特异性。     

六种石磺科贝类超氧化物歧化酶和酯酶同工酶分析

采用聚丙烯酰胺垂直板凝胶电泳分离技术对中国东南沿海的石磺科6种石磺的腹足、肝胰脏两种组织的超氧化物歧化酶和酯酶同工酶进行分析.明确了其酶谱的特征及分布,并利用聚类分析方法对种间的亲缘关系进行了研究.同工酶聚类分析显示,紫色疣石磺(Peronia verruculata)和小紫疣石磺(Peronia sp.)的亲缘关系最近;里氏拟石磺(Paraoncidium reevesii)和白底拟石磺(Paraoncidium sp.)聚为一类;平疣桑椹石磺(Platevindex mortoni)和瘤背石磺(Onchidium struma)聚为一类.种间的个体酶谱表型有差异,同属的种间差异小于不同属的种间差异.酶谱的差异程度与形态分类学中的亲缘关系相近.利用超氧化物歧化酶同工酶和酯酶同工酶酶谱表型也可以作为一种蛋白分子标记应用于石磺科属种的分类鉴定.     

乳酸脱氢酶与酯酶同工酶同板染色法

介绍一种在同一块凝胶板上染乳酸脱氢酶(LDH)与酯酶(EST)的染色方法. 该同板染色法利用两种同工酶显色反应互不干扰和颜色不同的特点, 先染LDH, 后染EST, 可以在同一块胶板上得到两种同工酶清晰的酶带, 每一种酶的酶带与单板染色的酶带完全一样. 这种染色法, 能节省同工酶分析所需的试剂、时间和经费, 也便于样品的鉴定与比较, 是一种经济有效的方法. 此方法, 同样适用于苹果酸脱氢酶(MDH)与酯酶等同工酶的同板染色.     

两个猪苓菌株生长速度及酯酶同工酶比较研究*

对不同地理分布的猪苓纯培养菌株进行了种性和酯酶同工酶的比较研究,结果表明,鸡爪苓(Z)纯培养菌株和猪屎苓(ZJ)纯培养菌株的种性有很大不同,两个纯培养菌株的酯酶同工酶酶带类型差异较大,亲缘关系较远。     

三种蜘蛛酯酶同工酶的比较研究

应用聚丙烯酰胺凝胶电泳对狼蛛科的拟水狼蛛、蟹蛛科的三突花蛛和肖蛸科的鳞纹肖蛸3种蜘蛛的酯酶同工酶进行了比较研究。结果表明,不同科的蜘蛛酯酶同工酶种问差异性大并有着明显的种簇特异性,推测它们的酯酶同工酶酶谱中的区带组受不同的基因位点控制,且各自的基因位点数不等;同种蜘蛛的雌蛛和雄蛛之间也有各自的酯酶同工酶谱型,但差异小,其控制基因位点大体相同。这样,我们从分子的水平上讨论了酯酶同工酶的差异性可以用来作为识别物种的附加指标。     

家鸽不同组织同工酶的初步研究

利用聚丙烯酰胺凝胶电泳方法对家鸽不同组织的酯酶同工酶、乳酸脱氢酶同工酶的酶谱进行了观察和试验分析。计算了家鸽8种组织酯酶同工酶各自酶带的Rf值。显示出家鸽8种组织的酯酶同工酶酶带和7种组织的乳酸脱氢酶同工酶酶谱分布特征具有明显的组织特异性。初步分析了基因位点与酶谱带型之间的相互关系,对于鸟类的生化分析和遗传研究有参考作用。     

6-BA对平菇和香菇菌丝体两种同工酶的影响

通过在平菇、香菇的马铃薯液体培养基中添加不同浓度的 6 BA(6 苄基腺嘌呤 ) ,应用聚丙烯酰胺凝胶垂直平板电泳技术 ,探讨了 6 BA对平菇、香菇菌丝体酯酶 (EST)和过氧化物酶 (PER) 2种同工酶的影响。结果显示 ,6 BA浓度在 5 g/L培养液和 15 g/L培养液时分别诱导出平菇、香菇菌丝体中各 1条新的酯酶同工酶带产生 ,不同的 6 BA浓度对平菇、香菇菌丝体其余的酯酶同工酶带强度也有影响 ;6 BA不能诱导平菇和香菇菌丝体中新的过氧化物酶同工酶产生 ,但在浓度为 15 g/L培养液时可使PER同工酶带增强 ;6 BA对平菇和香菇菌丝体中EST ,PER2种同工酶的Rf值没有影响。     

Identification and development of a genetically closely-linked carboxylesterase family of the mouse liver

Six carboxylesterase isozymes (viz. ES-1, ES-6, ES-9, ES-20, ES-22 and ES-24), governed by esterase gene cluster 1 on chromosome 8 of the house mouse, were identified electrophoretically in liver supernatants using their biochemical, genetic and developmental characteristics. ES-1 and ES-20 were expressed as liver-specific forms. The peri- and postnatal development of the six isozymes indicated that they were individually regulated at the genetic level, although the isozymes were regulated as a group when compared to genetically unrelated esterases. The concept of evolutionary divergence following repeated gene duplication of an ancestral esterase structural gene was extended to cover divergence of the temporal (regulatory) genes associated with the multigene family. Allelic variation of the temporal genes was more limited than that of the corresponding structural genes.     

Geographic Variation of Esterase Isozymes in Populations of Alternaria mali on Apple Leaves

The 500 monoconidial isolates of Alternaria mali occurring in different locations, Suweon, Cheongju, Kochang, Daegu and Jinju, Korea, in 1983 were used to examine geographic variation of esterase isozymes. The electrophoretic patterns of esterases were qualitatively and quantitatively different between isolates. The 14 different bands were detected on the basis of the decreasing electrophoretical mobility, although all bands were not present in any of the isolates. A comparison of the frequency of esterase isozymes at different bands showed marked variations among the geographic locations. The geographic distance between A. mali populations did not correlate strongly with divergence in esterase isozymes, whereas A. mali populations within a geographic feature were more closely related than populations separated by a mountain range.     

Purification and characterization of esterases D1 and D2 from human erythrocytes. Evidence that they are monomers

Esterase D1 and esterase D2, two common esterase D (EC 3.1.1.1) isozymes, were isolated and purified from human erythrocytes. Their substrate specificity, pH profile and Km values were essentially identical. Their molecular mass was the same at 34 kDa on sodium dodecyl sulfate/polyacrylamide electrophoresis and at 27 kDa on Sephadex G-100 gel filtration. Antisera to each of the esterase D1 and esterase D2 isozymes were successfully raised in chickens; each antiserum reacted identically with both isozymes. These findings indicate that the isozymes are close to each other in structure. The fact that the molecular mass of the esterase D1 and esterase D2 isozymes computed on sodium dodecyl sulfate/polyacrylamide electrophoresis was close to that obtained on Sephadex G-100 gel filtration in non-dissociating buffer indicates that the isozymes are not dimers bound by disulfide bonds or a noncovalent force. These facts together indicate that the esterase D isozymes are monomers, contrary to the prevailing view that they are dimers.     

Developmental genetics of the esterase isozymes of Fundulus heteroclitus

The ontogeny of the esterase isozymes of the teleost, Fundulus heteroclitus, has been investigated. One group of esterase isozymes is present at all stages of development, whereas other esterase isozymes only very gradually appear at later stages of development, or abruptly appear at such dramatic developmental events as hatching. The ontogeny of these isozyme patterns is interpreted as the expression of differential regulation of separate esterase genes. The general pattern of teleost esterase gene activation is similar to that reported for birds and mammals. Allelic variation was detected at two of the esterase loci. On the basis of electrophoretic mobility, substrate specificity, inhibitor specificity, genetic variation, and ontogeny of esterases, there appear to be at least 15 different esterase isozymes, which constitute 6–8 groups, each of which is probably encoded in one or more genetic loci.This study was supported by NSF Grant GB 544OX to Professor C. L. Markert and an NSF Graduate Fellowship to G. S. Whitt.     

普通伏翼不同组织3种同工酶的比较

采用聚丙烯酰胺不连续凝胶垂直板活性电泳,分析了普通伏翼的心、肝、肾、胸肌、肺、胃、小肠、舌8种组织的酯酶(EST)、乳酸脱氢酶(LDH)、苹果酸脱氢酶(MDH)3种同工酶。结果表明:普通伏翼8种组织的3种同工酶存在差异,其分布具有明显的组织特异性,与器官或组织所执行的功能有关。     

盾叶薯蓣自然变异类型间的比较研究

对取自云南期纳的二倍体和三倍体盾叶薯蓣植株以及重庆城口的四倍体植株进行了形态学、染色体数目及同工酶的比较研究。结果表明,这3种类型植株在叶片形态上有明显的差异。二倍体的染色体数目是2n=2x=20,三倍体为2n=3x=30,四倍体为2n=4x=40;酯酶(EST)和超氧化物歧化酶(SOD)酶谱显示,3种类型植株之间具有明显的相关性,但多倍体较二倍体的酶带条数少且着色浅。这些差异除了有地理分布不同的因素外,倍性的不同也是很重要的因素。     

Isoenzyme Polymorphism in Flowering Plants

Esterases, leucine aminopeptidases, catalases and acid phosphatases from pollen of Oenothera organensis were studied electrophoretically. A study was made using several different extraction media to see the effect it had on esterase migration and stainability. It was found that different extraction media resulted in significant changes in migration rates and stainabilities of the esterase isozymes. Anodal esterases and leucine aminopeptidases from 13 inbred lines of maize were studied and are reported. Preliminary genetic studies of two of the esterase isozymes from maize pollen are reported. A survey of anodal esterases and leucine aminopeptidases from the pollen of 11 genera of diverse angiosperms is presented.     

Isozyme variations inAegilops andTriticum. III. Chromosomal basis of the esterase isozyme production in different organs of Chinese spring wheat

Developmental changes of esterase isozymes from the germination to the heading stage of normal and aneuploid lines of common wheat,Triticum aestivum cv. Chinese Spring were studied. A total of twenty major isozymes (Bands 1E to 20E) were observed, some of which were further separated to two to three closely located bands. Among these bands, 1E, 2E, 3E, 5E, 7E, 11E, 14E and 16E were found to be leaf-specific isozymes and 9E, 10E, 13E, 15E, 17E and 18E were seed-specific. Leaf-specific isozyme bands 1E, 2E and 5E are controlled by genes on three homoeologous chromosomes group 6, leaf-specific bands 7E, 11E, 14E and 16E and seed-specific bands 9E, 10E, 13E, 15E, 17E and 18E are under control of genes on homoeologous chromosomes of group 3. On the other hand, two bands, 19′E and 19″E are controlled by genes on chromosomes of homoeologous group 2 in roots of seedlings 10 days old. The present investigation showed that the genes for esterase production located on chromosome 6B had large effects in mature leaves, but chromosomes 6A and 6D had little effect on the esterase isozymes in homoeologous group 6. Genes located on chromosomes 3A, 3B and 3D have a large function in germinating seed; however, chromosomes 3B had little effect on the esterase isozymes in the mature leaf. Present findings confirmed that the chromosomes of the A, B and D genomes have different functions in the production of proteins or enzymes. Contribution from the Laboratory of Genetics, Faculty of Agriculture, Kyoto University, Japan, No. 401.     

Biochemical characterization and purification of esterases from three strains of German cockroach,Blattella germanica (Dictyoptera: Blattellidae)

Three strains of German cockroach, Blattella germanica (L.) showed varying levels of resistance to chlorpyrifos, methyl parathion, propoxur, bendiocarb, and cypermethrin. The general esterase activity was at least twofold higher than susceptible strain. The subcellular distribution studies revealed that the majority of the esterase activity is present in the 100,000g cytosolic fraction. Only a small portion of the activity was membrane bound. Using non-denaturing gel electrophoresis, ten isozymes were identified in German cockroaches. These isozymes were isolated individually from the gels and analyzed for differences in activity. The isozymes E5, E6, and E7 of resistant strains had significantly higher specific activities when compared with the susceptible strain. The purification process using various column chromatography and preparative gel electrophoresis resulted in 9–11% of total esterase recovery. About double the amount of E6 was recovered from the resistant strains when compared with the susceptible strain. Kinetic analyses of E6 did not indicate differences in K_m and V_max values between the resistant and susceptible strains. Also, inhibition of esterase activity by paraoxon, chlorpyrifos, and propoxur did not suggest any structural differences in esterase E6 between strains. The results suggest that the increased production of E6 esterase contributes to insecticide resistance in German cockroaches. The role of E6 may be sequestration of toxic molecules rather than hydrolysis. © 1996 Wiley-Liss, Inc.     

Electrophoretic Pattern of Larval Esterases in Field and Laboratory-selected Strains of the Tobacco Cutworm,Spodoptera litura (Fabricius)

This research was performed to find out and characterize the specific esterase isozymes related to OP and pyrethroid resistance in the tobacco cutworm Spodoptera litura (Fabricius). Two laboratory strains (DSR₅ and CSR₄) of S. litura, selected with deltamethrin and chlorpyrifos-methyl, had higher larval esterase activities than Hamancollected (HC) strain. Ten esterase isozymes were separated in the wild HC strain on 6.5% nondenaturing polyacrylamide gel electrophoresis (pH 8.3), but only 5 of them were detected in the two strains selected with insecticides. The isozymes were designated from E1 to E10 according to their mobility to cathode. E4 was stained more strongly in both laboratory-selected strains than in HC strain. The frequency of E2 in DSR₅ strain was higher than in HC strain. E2 and E4 were proved to be arylesterase and carboxyesterase, respectively, according to enzyme inhibition tests. Thus decreased number and increased intensity of esterase bands in DSR₅ and CSR₄ strain may be associated with insecticide resistance, resulting from selections successively with insecticides for several generations.