利用玉米种子白蛋白和球蛋白乳酸聚丙烯酰胺电泳鉴定品种( 英文)

采用改进的乳酸 - PAGE技术对不同玉米 (Zea mays)自交系和杂交种种子的白蛋白和球蛋白进行了系统电泳分析 ,发现此方法具有高度的分辨力和稳定性。把电泳图依其迁移率分为 α、β、γ和 ω4个带区。各自交系和杂交种的电泳图均有其鲜明特征 ,彼此可以区别 ,视作它们各自的“指纹”。全籽粒电泳谱带与胚的电泳谱带基本相同。胚乳产生谱带少而弱 ,且大都和胚的谱带发生重叠。亲本自交系的不同谱带在杂种 F1 发生互补。杂种谱带等于两亲本共同谱带与各自独有谱带之和。而且正、反交 F1 谱带以及它们两亲本种子提取液机械混合产生的谱带完全相同 ,表明谱带的差异是一种由核基因控制的遗传性状。F1 的谱带类型可由两亲本谱带类型预测 ,也可由两亲本种子提取液混合而产生的电泳图所演示     

正常与感染粘孢子虫鲫鱼主要组织可溶性蛋白质电泳比较

采用SDS-聚丙烯酰胺凝胶电泳技术,对正常和感染粘孢子虫的鲫鱼动脉球、肝脏、鳃、脑、肠、肌肉、眼、鳍8种组织可溶性蛋白质进行分析比较。结果表明：病鱼的动脉球、肝脏、鳃、脑、肠5种组织的蛋白质电泳谱带与正常鱼相比有较大变化,动脉球缺失1条谱带同时又诱导出1条新谱带;肝脏有9条谱带缺失;鳃有2条谱带缺失的同时诱导出3条新谱带;脑中新增3条谱带;肠中有5条谱带缺失同时产生3条新谱带。这些变化可作为辅助疾病诊断的生化指标,为从生化水平上揭示该病的致病机理及开展有效的早期防治研究奠定基础。     

磁处理水对鲫鱼血清蛋白和乳酸脱氢酶同工酶的影响

本试验采用聚丙烯酰胺凝胶电泳技术,比较分析了经磁处理水饲养的鲫鱼血清蛋白谱带和ＬＤＨ同工酶谱带。结果表明：其血清蛋白和乳酸脱氢酶（ＬＤＨ）同工酶的电泳谱带与对照相比有明显差异。血清蛋白谱带平均变化率为３２．５％,ＬＤＨ同工酶谱带平均变化率为１０％。两种谱带均以饲养１０ｄ的变化率最高。结果提示了磁处理水对鲫鱼最佳影响时期。     

RAPD标记在山葡萄种质鉴定中的应用

采用修改后的CTAB 法获得了高质量的基因组DNA 。利用RAPD 标记技术对山葡萄7 份种质进行鉴定, 用4 个引物(从30 个引物中筛选)对试材进行PCR 扩增, 共扩增出30 条谱带, 平均每条引物产生7.5 条谱带, 其中21 条谱带为多态性谱带, 占总谱带数的70%。不同引物扩增的谱带数不同, 范围在6~9 条之间。利用4 个引物扩增出的多态性谱带可以将7 份山葡萄种质区分。     

部分美国小麦种质资源醇溶蛋白遗传多样性分析及其亚基对品质性状的影响

为了解67份美国材料的遗传多样性及其醇溶蛋白亚基对品质性状的影响,利用酸性聚丙烯酰胺凝胶电泳(APAGE)技术进行醇溶蛋白谱带分析,测定了面团流变学特性及理化品质。结果表明,在67份美国材料中共分离出1332条谱带,49种不同迁移率类型的谱带,大部分谱带具有多态性。单个材料谱带总数变异幅度为13~28。谱带数在α、β、γ、ω4个区的分布存在较大差异。没有发现电泳谱带完全相同的材料。GS值变异范围0.54~0.90,平均值为0.731。在GS=0.607水平上,聚类分析将这67份材料分为6类。49条不同迁移率的谱带中有17条谱带与36项品质性状的相关性达到显著或极显著差异。6条谱带(迁移率为49.6、56.2、56.7、62.2、79.4、86.8)与湿面筋含量、蛋白质含量和沉淀值呈正相关,而迁移率为60.5的谱带与之呈负相关。11条谱带(迁移率为26.5、42.0、49.6、52.5、56.2、56.7、62.2、64.1、72.0、79.4、86.8)与面团稳定时间、面团形成时间、延伸面积等面团流变学特征呈正相关,而迁移率为34.4、47.5、49.0、60.5、69.4、85.4的6条谱带则与之呈负相关。说明供试材料间存在着丰富的遗传多样性以及与优质品质相关的谱带,为进一步利用这67份种质资源和优质小麦品种的选育提供了理论依据。     

两种纤毛虫营养细胞和休眠细胞蛋白组成的比较分析

应用生化抽提和SDS-PAGE方法显示,膜状急纤虫(Tachysoma pellionella)的营养细胞含38条蛋白谱带,休眠细胞含29条蛋白谱带,两者共有谱带为26条,特有谱带各为12条和3条,相似度为77.6%;休眠细胞的包囊壁含22条蛋白谱带,细胞脱包囊后残留的包囊壁含15条蛋白谱带,两者共有谱带为14条,特有谱带各为8条和1条,相似度为76%。包囊游仆虫(Euplotes encysticus)的营养细胞和休眠细胞均含23条蛋白谱带,两者共有谱带为19条,特有谱带各为4条,相似度为82.6%;休眠细胞的包囊壁和细胞脱包囊后残留的包囊壁均含20条蛋白谱带,两者共有谱带为19条,特有谱带均为1条,相似度为95%。结果表明,两种纤毛虫的营养细胞向休眠细胞转化过程中,细胞结构的主要蛋白质成分发生了明显变化,这些变化与细胞在不同生理状态下结构的分化及其生命活动特征相关。形成“毛基体吸收型包囊”的急纤虫与形成“毛基体非吸收型包囊”的游仆虫相比较,前者营养期和休眠期细胞蛋白组成有更明显的差异,这可能与其细胞结构更大程度的脱分化有关;根据纤毛虫休眠细胞的包囊壁和细胞脱包囊后残留的包囊壁两者蛋白组成的差异推测,前者包囊壁可能含有与休眠细胞生命活动相联系的“活性”成分。     

特种稻谷蛋白亚基的研究

以陕西红香米、绿香米、紫糯、茉莉香米、黑帅、黑宝为实验材料,采用SDS-PAGE方法在pH为8.9,电流为15mA条件下,对其种子的谷蛋白进行了电泳分析,寻找其特征及其之间的差异。结果表明绿香米、黑帅都具有9条特征谱带;茉莉香米具有7条特征谱带;紫糯具有6条特征谱带;黑宝具有5条特征谱带;红香米具有4条特征谱带。分子量较高、蛋白谱带较多的品系为绿香米和黑帅(6～5条);分子量较高、蛋白谱带较少的品系为茉莉香米(2条)。     

半夏种内不同居群酯酶和超氧化物歧化酶同工酶酶谱特征分析

对栽培在同一生境下的16个半夏〔Pinellia ternata（Thunb.）Breit.〕居群同一生长期叶片中酯酶（EST）和超氧化物歧化酶（SOD）同工酶酶谱进行了比较分析,结果表明：EST和SOD同工酶酶谱在各居群间,甚至在同一居群内除少数共有的特征谱带外存在着较为明显的频率差异。EST同工酶在半夏各居群中最多可显示12条谱带,其中在慢区的第5（弱带）和快区的第10（强带）2条谱带为各居群所共有的特征谱带;SOD同工酶在半夏各居群中最多显示9条谱带,其中在慢区的第2（弱带）、快区的第5（强带）和第9（强带）3条谱带为各居群所共有的特 征谱带。     

5种樱桃属植物的POD、CAT和SOD同工酶分析

采用聚丙烯酰胺垂直平板凝胶电泳技术对5种樱桃属植物的过氧化物酶(POD)同工酶、过氧化氢酶(CAT)同工酶和超氧化物歧化酶(SOD)同工酶的酶谱特征进行分析,结果表明:5种樱桃属植物共电泳出12条POD同工酶酶带、4条CAT同工酶酶带和8条SOD同工酶酶带;其中,POD同工酶酶谱具有5条共同谱带,CAT同工酶4条,SOD同工酶5条。冬季休眠期的樱桃属植物,POD与SOD同工酶谱带的多样性比较丰富,不同植物之间的谱带差异较大;而CAT同工酶谱带差异不明显。     

7个品种变叶木过氧化物酶同工酶分析

本试验对7个品种变叶木的形态特征进行比较,并利用聚丙烯酰胺凝胶电泳,对各品种过氧化物酶同工酶进行分析。结果表明,各品种的过氧化物酶同工酶谱不一;7个品种变叶木共有14条谱带,2号谱带最多为11条,5号谱带最少为6条;其谱带分为3个区(A=0~0.18,B=0.18~0.35,C=0.35~0.5);并对7个品种的亲缘关系进行聚类分析。     

Histochemical localization of lactate dehydrogenase isoenzymes in human skeletal muscle fibers.

Lactate dehydrogenase (LDH) activity was histochemically localized in fibers of the vastus lateralis muscle of men and for comparative purpose in the soleus and plantaris muscleo of rats. Human muscle fibers were identified as fast twitch (FT) or slow twitch (ST) from the histochemical stain for myofibrillar adenosine triphosphatase activity. Rat skeletal muscle fibers were classified as fast-twitch-oxidative-glycolytic (FOG), fast-twitch-glycolytic (FG), or slow-twitch-oxidative (SO) on the basis of NADH-diaphorase and myofibrillar adenosine triphosphatase activities. Heart-type (H) LDH was identified by inhibition of the muscle-type (M) isozyme with 4 M urea. Total LDH as estimated histochemically was highest in the human FT and rat FG fibers. This was predominantly the M-LDH isozyme. ST fibers of human and SO fibers of rat skeletal muscle had the least total LDH but the most H-LDH activity. The FOG fibers of rat muscle contained a total LDH activity intermediate to that of the FG and SO fibers and a combination of H- and M-LDH. There were no fibers in the human muscle samples studied that had LDH activities similar to the FOG fibers of rat muscle.     

Patterns of gene expression during teleost embryogenesis: Lactate dehydrogenase isozyme ontogeny in the medaka (Oryzias iatipes)

The ontogeny of the lactate dehydrogenase (LDH; EC 1.1.1.27) isozymes during medaka (Oryzias latipes) embryogenesis was determined after the genetic and molecular bases of this multilocus isozyme system were established. Three LDH loci are differentially expressed among the tissues of the adult medaka. The LDH-A locus was expressed almost exclusively in the white skeletal muscle, the LDH-B locus in all tissues examined, and the LDH-C locus in the eye and brain. The contribution of each of these LDH loci was quantitatively determined throughout early medaka embryogenesis by using a combination of electrophoretic, immunochemical, and spectrophotometric procedures. LDH-B₄ is present throughout embryogenesis and is the predominant LDH isozyme during this period. LDH-C subunit activity was first detected 146 hr after fertilization (26°C), 142 hr prior to hatching. LDH-A subunit activity, however, was not detected until after hatching and, then, only as heterotetramers containing LDH-B subunits. The pattern of LDH gene expression during medaka embryogenesis was compared with the patterns of LDH gene expression during early development in five other teleost species. Some common patterns of differential LDH gene expression appear to exist among the teleosts. In all species examined, isozymes encoded in at least one LDH locus, A and/or B, were present throughout development. Those isozymes present continually during embryogenesis also tend to be active in a wide variety of differentiated tissues in the adult fish. Conversely, LDH isozymes which are active in a restricted number of adult tissues are detected only later in embryogenesis. The initiation of LDH-C gene expression, however, is closely coupled with morphological and functional differentiation of those cells in which this locus is predominantly expressed in the adult.     

Localization of lactate dehydrogenase isozymes in human muscle tissues by the mixed aggregation immunocytochemical technique.

Lactate dehydrogenase (LDH) isozyme composition and localization was determined in sections of skeletal, heart and smooth muscle by the mixed aggregation immunocytochemical method using first antibody directed against purified human LDH-A4 (M4) or LDH-B4 (H4) followed by the enzymes LDH-A4 and LDH-B4, respectively. An even distribution of the two monomers in all fibres was seen with heart muscle and smooth muscle. Heart muscle had a low concentration of A-monomers and a high concentration of B-monomers, whereas the smooth muscle had equal concentrations of the two monomers. In contrast, skeletal muscle from m. quadriceps femoris was found to be composed of two muscle fibre types, one containing mainly A-, the other mainly B-monomers. On the basis of succinate dehydrogenase activity it was shown that the red (type 1) fibres contain mainly B-monomers and the white (type 2) fibres mainly A-monomers of LDH.     

几种鲤科鱼类及杂种的乳酸脱氢酶同工酶的比较

用聚丙烯酰胺凝胶电泳法,分析了14种鲤科鱼类和4个杂交种的肌肉、心脏、晶状体、脑和肾脏的乳酸脱氢酶(LDH)同工酶。根据LDH酶谱的异同可将雅罗鱼亚科的5种鱼分成两组:草鱼、鳡和鳤;青鱼和赤眼鳟。鳊亚科中青梢红鲌的LDH谱形与长春鳊、团头鲂和红鳍鲌相似,而与(歺又鱼)条有明显的区别。鲢亚科中的鳙与鲢的谱形相似。密鲴亚科中的细鳞斜颌鲴与黄尾密鲴的谱形也非常相似。将谱形相似的鱼类进行杂交,如细鳞斜颌鲴♀×黄尾密鲴♂、鳙♀×鲢♂,不仅杂种的成活率高而且可繁殖后代。谱形相差很大的鱼类进行杂交,如草鱼♀×团头鲂♂及草鱼♀×鳙♂,杂种的成活率都很低。这些结果和酶谱所显示的关系是一致的。       

Genetic characterization of four wild species of Chinese marmots using microsatellite markers

Marmots are large ground squirrels, and 14 species have been reported in the world, including four species of marmots (Himalayan marmot, Tarbagan marmot, gray marmot and long-tailed marmot) living in China. Although these biological resources are abundant in China, information regarding their genetic features is lacking, hampering further study regarding them. The aims of this research were to evaluate genetic variations of four species of Chinese wild marmots, and analyzed kinship of these marmot populations. In the current study, we collected samples of four species of Chinese wild marmot and analyzed the effective allele number, gene diversity, the Shannon index, and polymorphism information to evaluate genetic variations using 13 microsatellite loci. Based on Nei’s genetic distance using the unweighted pair group method, we constructed a dendrogram to analyze the population kinship. We determined that all four Chinese marmot species had high genetic polymorphisms and departure from Hardy-Weinberg equilibrium. The Chinese marmots to be divided into two large groups: Himalayan marmot was independent group. Tarbagan marmot, gray marmot and long-tailed marmot were others; Tarbagan marmot and gray marmot showed a close kinship with each other, but long-tailed marmot did not have a close relationship with the other species. The high polymorphisms and the kinship of Chinese marmot populations were correlated with geographical terrain of their habitat. Himalayan marmot was characterized as living in unique alpine meadows in Qinghai-Tibet plateau and was affected by terrain; however, Tarbagan marmot, gray marmot and long-tailed marmot were characterized as living in grassland or alpine grassland and were not affected by terrain. Genetic features of Chinese wild marmots were investigated in this study. This may give using information regarding protection of Chinese wild marmot resource and further application of biomedical research.     

白鲢个体发育过程中同工酶基因的表达与调控研究

采用淀粉或聚丙烯酰胺凝胶电泳及特异性组织化学染色技术研究了白链早期发育过程中（从未受精卵到卵黄吸尽期）及成体不同组织（脑、眼、心、肌、肾、肝）中六种同工酶系统（ＬＤＨ,ＭＤＨ,ＩＤＨ,ＡＤＨ,ＳＤＨ,ＥＳＴ）的分化表达谱式,白链各同工酶基因的表达具有明显的组织特异性早期发育过程中,ＡＤＨ和ＳＤＨ基因均无活性,其它四种同工酶则具有不同的个体发育谱式。值得一提的是,由取自同一地区的两对白链的受精卵的形     

Lactate dehydrogenase isozymes in type I,IIA and IIB fibres of rabbit skeletal muscles

Summary Lactate dehydrogenase (LDH) isozyme patterns were analysed by polyacrylamide (PAA) slab gel electrophoresis in extracts prepared from various rabbit skeletal muscles of defined fibre composition and by PAA microelectrophoresis of microdissected, histochemically typed single muscle fibres. The results obtained by electrophoresis of whole muscle extracts generally agreed with the data obtained from single fibre electrophoresis, i.e. the LDH isozyme pattern corresponded to that of the predominant fibre type. Type I Fibres from soleus and semitendinosus muscles were characterized by a unique pattern of all 5 LDH isozymes with a predominance of LDH-1, 2 and 3. The major fraction (80%) of the type II fibres from extensor digitorum longus and tibialis anterior muscles contained only LDH-5 (M₄). About 20% of the type II fibres contained in addition to LDH-5 small amounts of LDH-4 and LDH-3. The fraction of fibres containing LDH-5, LDH-4, and LDH-3 was similar (ca. 20%) in the histochemically defined IIA and IIB subpopulations In view of the fact that the major fractions of rabbit IIB fibres display low and of IIA fibres high aerobic oxidative capacities (Reichmann and Pette 1982), these data indicate that the expression of the H-subunit of LDH is not correlated with the aerobic-oxidative capacity of the fibre. It also appears not to be correlated with the presence of different myosin isoforms in IIA and IIB fibres.     

Flow cytometry of human spermatozoa

Summary Lactate dehydrogenase (LDH) isozyme composition and localization was determined in sections of skeletal, heart and smooth muscle by the mixed aggregation immunocytochemical method using first antibody directed against purified human LDH-A₄ (M₄) or LDH-B₄ (H₄) followed by the enzymes LDH-A₄ and LDH-B₄, respectively. An even distribution of the two monomers in all fibres was seen with heart muscle and smooth muscle. Heart muscle had a low concentration of A-monomers and a high concentration of B-monomers, whereas the smooth muscle had equal concentrations of the two monomers. In contrast, skeletal muscle from m. quadriceps femoris was found to be composed of two muscle fibre types, one containing mainly A-, the other mainly B-monomers. On the basis of succinate dehydrogenase activity it was shown that the red (type 1) fibres contain mainly B-monomers and the white (type 2) fibres mainly A-monomers of LDH.     

Enzyme patterns in trisomy 19 of the mouse

Activity patterns of cytosolic and mitochondrial enzymes of carbohydrate and amino acid metabolism have been measured in murine trisomy 19. In spite of marked hypoplasia, no significant alterations of the patterns (per gram of organ weight) were observed, with the exception of glutamate oxaloacetate transaminase (GOT-1), and phosphoglycerate mutase (PGAM). Clear-cut gene dosage effects in liver, brain, heart, skeletal muscle, and erythrocytes of fetal and newborn mice, confirm the assignment of GOT-1 to chromosome 19. Data obtained for PGAM demonstrate that one of the two different subunits leading to organ-specific isozyme patterns of the dimer enzyme protein is coded on chromosome 19 (gene Pgam-1). Dosage effects are fully expressed in liver, brain, and erythrocytes (AA-type isozyme), but not in skeletal muscle (BB-type isozyme). Dosage effects on the hybrid AA-AB-BB-isozyme pattern in the course of development of the heart muscle, were demonstrated by means of quantitative activity measurement after electrophoretic separation. The comparison of enzyme patterns of eusomic and trisomic erythrocytes, produced after injection of fetal stem cells into irradiated adult carriers (transplantation chimaeras), revealed enzyme activity ratios that were similar to those produced by erythrocytes of adult euploid and trisomic mice. This is in agreement with the chromosome assignments and dosage effects mentioned above.     

Modulation of Lactate Dehydrogenase Isozymes by Modified Base Queuine

The modified base queuine is a nutrient factor for lower and higher eukaryotes except yeast. It is synthesized in eubacteria and inserted into the wobble position of specific tRNAs (tRNA_GUN) in exchange of guanine at position 34. The tRNAs of Q family are completely modified in terminally differentiated somatic cells. However, mainly free queuine is present in embryonic and fast proliferating cells, tRNA remains Q deficient. Lactate dehydrogenase (LDH) A mRNA and LDH A protein is known to increase when cells are grown in hypoxic conditions. In the present study, the level of LDH isozymes is analyzed in different tissues of normal and cancerous (DLA) mice and the effect of queuine treatment on LDH isozyme is observed. LDH A isozyme is shown to increase in serum and liver of DLA mice. The level and activity of LDH A decreases on queuine treatment. In skeletal muscle and heart, LDH A isozyme decreases while LDH B increases in DLA mice. Queuine administration leads to change back towards normal. In case of brain, LDH A increases but LDH B decreases in DLA mice. Queuine treatment leads to decrease in A4 anaerobic isozymes of LDH. The results suggest that queuine suppresses anaerobic glycolytic pathway, which leads to tumor suppression of DLA mice.