斯氏并殖吸虫乳酸脱氢酶,苹果酸脱氢酶及酯酶同工酶的研究

本文采用Disc-PAGE电泳,首次对我国独有的斯氏并殖吸虫(Paragonimus skrjabini Chen,1959)成虫、童虫、囊蚴的乳酸脱氢酶(以下简称LDH)、苹果酸脱氢酶(以下简称MDH)和酯酶(以下简称EST)同工酶进行了研究。 在成虫、童虫、囊蚴间,LDH、MDH、EST同工酶在酶带数、排列型式、Rf值、相对活性和优势酶带的位置都存在差异。 根据虫体和宿主组织同工酶谱的不同,可以认为是本虫本身所具有。 同工酶作为其分类指标时,不仅要比较不同虫种成虫稳定的同工酶谱,也要比较同工酶在个体发育型式间的差异。     

二倍体型与三倍体型卫氏并殖吸虫同工酶谱的发育变异及其分类学意义

采用生趣板状聚丙烯酰胺胶是民泳法分析比较了二倍体型与三倍体型卫氏并殖吸虫童虫、成虫的醛缩酶、已糖激酶和葡萄糖－６－磷酸脱氢酶的同工酶谱,结果显示２ｎ型与３ｎ型卫氏并殖吸虫在不同发育期之间及两型吸虫相应各发育期之间ＡＬＤ、ＨＫ及Ｇ６ＰＤ同工酶谱均显著变异,变异表现为两种方式,一种是在同一个基因座位区的同工酶的酶带数目     

异精效应在雌核发育彭泽鲫胚胎发育中的同工酶证据

采用聚丙烯酰胺凝胶垂直板电泳技术 ,对同源和异源精子激发雌核发育彭泽鲫子代胚胎发育过程中四种同工酶 (EST ,LDH ,MDH ,ME)的表达情况进行了比较研究。结果表明同源和异源精子激发的胚胎在MDH和LDH同工酶表达上存在明显差异。MDH同工酶的差异主要表现为 :在孵出期 ,异源精子激发的胚胎比同源精子激发的胚胎多出两条谱带MDH4’和MDH5’ ;LDH同工酶的差异表现为 :在原肠中期至肌肉收缩期的五个时期中 ,异源精子激发的胚胎比同源精子激发的胚胎多出 4条谱带 (LDH9’— 12’)。这种差异说明同源与异源精子对子代胚胎发育过程中同工酶表达的影响不同 ,可能属于“异精生物学效应”的一种表现形式     

黄颡鱼不同组织中同工酶的表达模式

采用聚丙烯酰胺凝胶电泳(PAGE)及特异性组织化学染色技术,研究了黄颡鱼成体眼、心脏、肾脏、肝脏、肌肉及尾鳍等6种组织中的4种同工酶(LDH、MDH、SOD、EST)分化表达模式。结果表明,黄颡鱼的同工酶EST、SOD、MDH具有明显的组织特异性,LDH无明显的组织差异,并对同工酶的组织表达特异性的生理意义进行分析和讨论。     

四类不同倍性鲫鱼在胚胎发育期间同工酶基因表达的比较分析

用聚丙烯酰胺梯度凝胶电泳比较分析了单倍体、二倍体、三倍体和复合四倍体4类不同倍性鲫鱼以及单倍体和二倍体鲤鱼在胚胎发育时期4种同工酶(EST,LDH,MDH,SOD)酶谱。结果表明,单倍体鲫鱼和单倍体鲤鱼胚胎与各自的二倍体胚胎相比,同工酶酶谱看不出差异;天然三倍体银鲫胚胎的MDH和SOD同工酶酶谱与二倍体鲫相似,但EST和LDH同工酶比二倍体增多了酶带,有的酶带如EST5和EST6还可在鲤鱼胚胎中找到相应的表达产物,提供了天然雌核发育三倍体银鲫杂交起源的证据;复合四倍体由于含有鲤鱼的一个外来基因组,其胚胎的基因表达有些与杂种类似,在所分析的4种同工酶酶谱中,都可观察到来自鲤鱼基因的影响。此外,在由源于不同复合四倍体个体的卵子发育形成的胚胎间,还观察到同工酶基因表达的异质性。     

高背银鲫、大口胭脂鱼及胭脂鲫(F1)不同组织的同工酶的表型分析

本文对高背银鲫、大口胭脂鱼及胭脂鲫（F1）不同组织的脂酶（EST）、乳酸脱氢酶（LDH）、苹果酸脱氢酶（MDH）、超氧化物歧化酶（SOD）四种酶的同工酶的表型进行了分析,结果表明：在肝脏中胭脂鲫EST多一条谱带,而在性腺中,LDH、MDH、SOD同工酶谱带,胭脂鲫与父母本均存在一定的差异。     

卫氏并殖吸虫染色体的研究近况

并殖属为吸虫纲中较复杂的一个属。全世界已报道四十多种,在我国有二十多种。仅依赖传统的分类方法对并殖吸虫分类似有一定困难。近年来,人们对并殖吸虫的染色体进行较广泛的研究。本文仅对卫氏并殖吸虫染色体及其在分类中作用的研究进展,简述于后。一、卫氏并殖吸虫的染色体组型按卫氏并殖吸虫的染色体数目可分二倍体型2n=22(n=11)和三倍体型3n=33(n=11)。按Levan等的方法进行染色体分类,二倍体型由一对(第一对大的中部着丝点染色体,四对(第2—5对)中等大的亚端部着丝点染色体,六对(第6—11对)小的中部、亚中部和亚端部着丝点染色体组成。…     

罗氏沼虾个体发育早期的同工酶研究

采用聚两燃酰胺梯度凝胶电泳技术,对罗氏沼虾个体发育早期9个时期的八种同工酶系统（EST、ALP、AMY、GDH、MDH、LDH、SOD、ME)进行研究,结果表明：SOD、ME在早期发育过程中酶谱相对稳定,SOD表现为三条谱带,ME表现为两条谱带;面EST、ALP、AMY、GDH、MDH、LDH则随发育其酶谱表现出明显差异,酶谱渐趋复杂。     

暗纹东方TUN 同工酶生化表现型的研究

利用聚丙烯酰胺凝胶垂直平板电泳方法,对暗纹东方|TUN|的心、肝、肾、肌、性腺5种不同组织的7种同工酶(EST、LDH、POD、MDH、SOD、SDH、α-AMY)进行了研究, 讨论了各同工酶的基因表达谱式,观察到EST同工酶存在着多态现象;LDH同工酶有二个基因位点,但只表现3条带,A与B亚基的结合受阻; MDH同工酶存在性别差异,说明决定MDH同工酶表达的因素在不同性别中存在差异;SOD同工酶有3个基因位点。各同工酶酶谱稳定,有组织特异性,但EST、MDH、SOD、POD同工酶在各组织器官中又表现出较大的一致性,有利于物种的鉴定。α-AMY与SDH只在个别组织中有活性,可能与特定组织与器官的形态发生与机能分化有关。 Abstract:By means of polyacrylamid gel electrophoresis seven isozymes(EST.LDH,SOD,MDH,SDH, α-AMY)in heart,liver,kidney,muscle and gonad of Fugu obscurus were studied.The gene expression patterns of each isozyme were analyzed.The results indicated that polymorphism was detected in EST isozymes;LDH isozymes had two loci,but only three bands could be observed,the random association of two subunits(A and B)were restricted;Some MDH isozymes existed sexual differences in identical tissues.The suggested that the factors controlling the expression of MDH isozymes were different between sexes.All the isozyme phenotypes exhibited tissue-specificity and stability,but EST,MDH,SOD and POD isozymes showed relatively consistence in the five tissues.Their characteristic bands could be used in species determination.The activities of SDH andα-AMY isozymes could only be detected in some tissues and closely correlated to the morphological or functional differentiation of those tissues or organs.     

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.     

四类不同倍性鲫鱼在胚胎发育期间同工酶基因表达的比较分析

Isozyme zymograms of esterase (EST), lactate dehydrogenase (LDH), malate dehydrogenase (MDH) and superoxide dismutase (SOD) were analysed by polyacrylamide gradient gel electrophoresis at different developmental stages of embryogenesis in 4 types of various ploidy crucian carp embryos, including haploids, diploids, natural triploids, and multiple tetraploids, and 2 types of haploid and diploid common carp embryos. Haploid embryos of crucian carp (Carassius auratus) and common carp (Cyprinus carpio) were produced by treating eggs with UV-irradiated milt from blunt snout bream (Megalobrama amblycephala). Natural triploid embryos were obtained from the eggs of gynogenetic silver crucian carp (Carassius auratus gibelio) inseminated with milt from red common carp. Multiple tetraploid embryos were also produced by gynogenesis from eggs of the newly discovered multiple tetraploid females inseminated with milt from red common carp. Gradient gel electrophoresis indicated that the band types and staining intensity of 4 isozymes expressed in haploid embryos of crucian carp and red common carp were similar to that in the correlative diploid embryos. In natural triploid silver crucian carp embryos, the zymograms of MDH and SOD isozymes were identical with that of diploid crucian carp embryos, but the EST and LDH isozymes manifested more new enzyme bands in comparison with diploid embryos. The corresponding expressed products of some bands in the triploid embryos, such as EST5 and EST6, could be observed also in red common carp embryos, which provided evidence for hybrid origin about the gynogenetic fish. The multiple tetraploids incorporated one foreign genome of red common carp, therefore, the effects of genes from the foreign genome could be observed in the multiple tetraploid embryos. Gene expression of the isozymes in the tetraploid embryos was somewhat similar to that in hybrids. Owing to interaction of triploid silver crucian carp genomes and common carp haploid genome, some isozyme bands, such as EST5 and EST6, changed in quantity, and some bands increased, such as s-SOD1, s-SOD2, s-SOD3 and s-SOD4 in the tetraploid embryos. Moreover, the heterogeneity was revealed among embryos developed from gynogenetic eggs of 3 different multiple tetraploid individuals.     

金鱼同工酶的发生遗传学研究——Ⅲ.金鱼同工酶基因座位的加倍与多倍性

以鲫鱼和金鱼为材料,用葡萄糖-6-磷酸脱氢酶(G6PD)、乳酸脱氢酶(LDH)和苹果酸脱氢酶(MDH)同工酶体系作为基因标志,从检测同工酶的多重组合形式来研究基因的加倍与演化。对彩鲫与金鱼G6PD和LDH同工酶的分析结果表明,它们均具有与四倍体鱼类相应的谱带。因而说明了金鱼的G6PD和LDH同工酶基因座位的加倍与染色体的多倍性有关,为金鱼是四倍体的假说提供了证据。而对MDH同工酶的分析却得到了与二倍体鱼类相同的谱带数。这可能与加倍基因发生突变而不表达有关。     

Comparison of gene representation between diploid and triploid Paragonimus westermani by expressed sequence tag analyses

Expressed sequence tag (EST) analysis of the diploid and triploid Paragonimus westermani genes was done to have a rapid and informative outlook of the gene-expression profiles of the parasites. Totals of 506 and 505 ESTs were generated from the diploid and triploid P. westermani cDNA libraries. Based on the BLASTx search results of the diploid P. westermani ESTs, 308 (60.9%) matched significantly with formerly identified genes and 198 (39.1%) showed no significant homology in the GenBank database. A similar homology pattern was shown from the triploid EST BLASTx search results with 346 (68.5%) sharing homology with previously identified genes and 159 (31.5%) showing no significant homology. The EST data from both libraries were analyzed and grouped into 9 categories. Comparison of the 2 EST pools revealed high similarities among the categories of the significantly matched genes. Single genes matched repeatedly were also observed in the 2 EST data. Some genes were found that are not yet characterized in P. westermani; these genes were matched by both the diploid and triploid ESTs. Further study of these genes may provide us with more understanding on the parasite's biology and their specific functions in the 2 strains.     

Isozyme expression in F1 hybrids between carp and goldfish

Interspecific genetic differences in malate dehydrogenase (MDH), lactate dehydrogenase (LDH), superoxide dismutase (SOD), and esterase (EST) isozymes in carp (Cyprinus carpio) and goldfish (Carassius auratus) were used to examine the allelic expressions in the hybrid between these species. A unique liver SOD and muscle LDH phenotype unambiguously identifies all presumed hybrid individuals. There was no evidence of F₂ or backcross phenotypes in hybrid individuals. Liver MDH and EST phenotypes in hybrids show a preferential expression of goldfish isozymes. Variation in the levels of carp liver MDH isozymes may result from the polymorphism of a regulatory mutation affecting isozyme expression, leading to gene silencing after duplication.This work was supported through NSERC (Canada) grants to James P. Bogart and John F. Leatherland.     

C-banding analysis of six species of lung flukes, Paragonimus spp. (Trematoda: Platyhelminthes), from Japan and Korea

We examined C-banded karyotypes of six species of lung flukes from Japan and Korea; diploid and triploid Paragonimus westermani, P. miyazakii, P. ohirai, P. iloktsuenensis and P. sadoensis, with special reference to their karyotypic diversification. C-band analysis between the diploid and the triploid westermani revealed that two of three homologues of the triploid resembled those of the diploid in C-band pattern, while the remaining chromosome showed a different pattern from any species examined here. This karyological evidence indicates that the triploid is allotriploid probably induced by interspecific hybridization between the diploid westermani and an unknown species; we, therefore, suggest that the triploid westermani is an independent species and synonymous with P. pulmonalis (Miyazaki 1978). As the morphologically similar three species, ohirai, iloktsuenensis and sadoensis, had the same C-band polymorphism in chromosome No. 4, these species are classified as the local races of P. ohirai. Paragonimus miyazakii has one common C-band (5q) with the diploid westermani, but other bands (1q, 4q, 6q, 7p and 7q) are different. From these observations, the six species examined are phylogenetically divided into three groups: (1) westermani group containing diploid and triploid (= pulmonalis) species, (2) miyazakii and (3) ohirai including two geographic races, iloktsuenensis and sadoensis.     

泽蛙雌核单倍体几种同工酶基因表达的研究

本研究用聚丙烯酰胺凝胶垂直平板电泳,比较泽蛙雌核生殖单倍体和泽蛙二倍体在鳃盖闭合期的乳酸脱氢酶同工酶(LDH)、苹果酸脱氢酶同工酶(MDH)、葡萄糖-6-磷酸脱氢酶同工酶(G-6-PDH)和酯酶同工酶(EST)的表型。实验表明,单倍体的上述同工酶区带数和活力与同胚期的二倍体比较,差异甚大。由此,作者认为,泽蛙单倍体问工酶基因的表达异常是由于缺少另一套染色体基因的相互作用。