家兔肌酸激酶的分离纯化和测定

[目的]提取分离纯化部分可溶解的肌酸激酶,应用于检测各种病变包括肌肉萎缩和心肌梗塞发生时的血清肌酸激酶水平改变。[方法]先用液氮处理家兔内侧腓肠肌和心肌,经过初步分离纯化得到肌酸激酶的粗提液,然后利用DEAE-52阴离子交换层析进一步纯化酶液,收集其中的活性峰,并用SDS-聚丙烯酰胺凝胶电泳检测其纯度。[结果]证明肌酸激酶是一个二聚体酶,带有两个相同的近似43kDa的亚基,同时显示家兔心肌和骨骼肌的肌酸激酶与其他物种的肌酸激酶有很大的同族性。[结论]新鲜心肌肌酸激酶的活性与贮存时间较长的心肌肌酸激酶的活性有很大差异。肌酸激酶同工酶具有重要的生理功能和临床医学应用价值。     

Human phosphoglycerate kinase and inactivation of the X chromosome

The fibroblasts derived from the skin of a woman heterozygous for an X-linked deficiency of phosphoglycerate kinase represented a mosaic. Two of 22 clones with normal glucose-6-phosphate dehydrogenase activity and hypoxanthine(guanine) phosphoribosyltransferase activity had no phosphoglycerate kinase activity detected by electrophoresis. Because the loci for glucose-6-phosphate dehydrogeniase and hypoxanthine(guanine)phosphoribosyltransferase are already known to undergo inactivation and to be on the short arm of the X chromosome and the locus for phosphoglycerate kinase is on the long arm, these observations support the conclusion that the entire human X chromosome can be involved in X inactivation.     

青海藏雪鸡和高山雪鸡血清淀粉酶同工酶酶谱研究

[目的]研究青海藏雪鸡和高山雪鸡血清淀粉酶同工酶酶谱。[方法]采用聚丙烯酰胺凝胶垂直平板电泳法对10只藏雪鸡和10只高山雪鸡及20只家鸡的血清淀粉酶同工酶酶谱进行了研究。[结果]藏雪鸡和家鸡血清淀粉酶均有AMY1、AMY2、AMY33种同工酶,且藏雪鸡的AMY1同工酶为单态,家鸡存在AMY1AB和AMY1BB表型呈现多态性,而高山雪鸡的AMY1同工酶不显现酶活性;3种鸡的AMY2同工酶均存在多态性;AMY3同工酶均表现为单态。藏雪鸡和家鸡血清淀粉酶AMY1同工酶的A和B区带相对迁移率分别为25.42%和18.33%;AMY2同工酶的A、B和C区带的相对迁移率分别为16.44%、13.70%和10.96%。[结论]该研究结果补充和丰富了藏雪鸡和高山雪鸡血液基因储备,并为其资源保护和利用及繁殖育种等提供了理论依据。     

C-type natriuretic peptide stimulates chicken myoblast differentiation through NPRB/NPRC receptors and metabolism pathway

Skeletal muscle development is closely related with the amount of meat production and its quality in chickens. Natriuretic peptides (NPs) play an important role in myotube formation and fat oxidation of skeletal muscle in animals. The effect of C-type natriuretic peptide (CNP), an important member of the NPs, and its underlying molecular mechanisms in skeletal muscle are incompletely understood. Treatment of myoblasts with CNP led to enhanced proliferation/differentiation and significantly upregulated (P<0.05) mRNA expression of the CNP receptors natriuretic peptide receptor B (NPRB) and the clearance receptor C (NPRC). In cells exposed to CNP, 142 differentially expressed genes (84 up-regulation and 58 down-regulation) (P<0.05) were identified by RNA-sequencing compared with those in control cells. Sixteen genes were significantly enriched (P<0.05) in the metabolic pathway, and six of them (phospholipase C β4, phospholipase C β2, phosphoglycerate mutase 1, creatine kinase B, peroxiredoxin 6 and CD38) were closely related to skeletal muscle development and differentially expressed. In conclusion, CNP stimulated differentiation of myoblasts by upregulating expression of the NPRB and NPRC receptors and enriching key genes in the metabolic pathway.     

Creatine phosphokinase in thyroid: isoenzyme composition compared with other tissues

Considerable creatine phosphokinase activity was demonstrated in thyroid glands of several species. After starch gel electrophoresis, thyroid exhibited a creatine phosphokinase isoenzyme pattern different from that of muscle, heart, and brain. The isoenzyme repertory of various tissues and a hybrid enzyme produced from muscle and brain enzyme are consistent with the postulated dimeric structure of creatine phosphokinase.     

猪血清酶和氟烷敏感性与肌肉品质关系的研究

 本文用54头ES系和30头斯格猪对血清肌酸激酶[CK，EC#-（2，7，3，2）]、乳酸脱氢酶[LDH，EC#-（1，1，1，2）]和氟烷敏感性与肌肉品质的关系进行了研究。结果表明：常规和药物应激后采血时血清CK活性与宰后45分钟肉质指标存在显著性相关（P＜0.01），血清LDH#-5同工酶和肉质指标高度相关（P＜0.001）。并且血清1ogCK值和LDH#-5同工酶在轻度PSE和正常肌肉猪之间存在极显著差异（P＜0.01）；氟烷阳性猪宰后表现典型PSE肌肉（斯格猪n=3），氟烷阴性宰后虽然未发生典型PSE肌肉，但存在轻度PSE肌肉胴体。     

分布稻田的三种不同体色泥鳅组织LDH同工酶的研究

本文报道了应用聚丙烯酰胺凝胶垂直板电泳分析研究分布于稻田的三种不同体色泥鳅在心肌,骨骼肌,肝脏,眼,血清七种组织器官中ＬＤＨ同工酶的表达特征。电泳图谱表明,这些组织分布和活力均具有明显的组织特异性;在脑组织ＬＤＨ２活性最强,其它组织如血清,眼,卵巢,肝脏,心肌等组织中ＬＤＨ３活力最强,此外,在肝脏中存在由Ｌｄｈ－Ｃ编码的Ｃ带。     

昆明裂腹鱼不同组织乳酸脱氢酶同工酶研究

采用不连续聚丙烯酰胺凝胶电泳法,对昆明裂腹鱼的眼球、心脏、肾脏、性腺、脾脏、肝胰脏、脑、鳃和肌肉等9种组织的乳酸脱氢酶同工酶(LDH)进行了测定。结果表明:昆明裂腹鱼LDH共有15条酶带表现出特异性,在不同组织中检出的LDH酶带数量不同,以肝胰脏中最多,共检出9条;在酶活性方面,LDH在脑、鳃、肌肉和肾脏等组织中活性最强,而在眼球组织中最弱。     

琅琊鸡IGF-Ⅰ基因多态性与肉用性的相关性

采用PCR-RFLP技术对琅琊鸡胰岛素样生长因子-Ⅰ(IGF-Ⅰ)5′非编码区PstⅠ酶切位点的多态性与生长发育的相关性进行分析.结果表明,琅琊鸡该位点存在2种等位基因(A和B),基因频率分别为61.7%和38.3%,其中A等位基因是有利基因.AA型个体活重、半净膛重和脂肪含量极显著高于BB型(P<0.01),其屠体重、全净膛重、腿肌重和胸肌重显著大于AB型和BB型(P<0.05),其剪切力和失水率也高于BB型(P<0.05,P<0.01).结果表明:等位基因A对琅琊鸡的肌肉产量和品质具有正效应.     

苹果叶片感染褐斑病过程中过氧化物酶同工酶变化的研究

 应用聚丙烯酰胺凝胶电泳,测定抗性不同的3个苹果品种过氧化物酶同工酶在接种褐斑病菌(Marssonina coronaria)后的变化规律。结果表明:苹果过氧化物酶同工酶在整个病程中呈类似常态曲线变化,症状出现时酶活性达最高峰,品种越感病,变化越强烈,表现在接种初期第12酶带增加早、增加量大;发病后期酶带数目比抗病品种减少更多。上述性状对病情诊断和幼苗抗性鉴定有参考价值。     

超重力处理对绿豆EST、POD和SOD同工酶的影响

采用垂直平板聚丙烯酰胺凝胶电泳技术,研究了绿豆在不同超重力处理下的EST同工酶、POD同工酶和SOD同工酶的变化情况。结果表明,在不同超重力处理条件下,绿豆的EST同工酶酶带数和活性变化较明显,增加了2条补偿性酶带,缺失了1条活性酶带,而POD、SOD同工酶超重力处理与对照无明显变化。只是萌发过的种子与干种子相比EST同工酶1条弱带增强;POD同工酶活性增强;SOD同工酶活性减弱。通过以上同工酶分析,证明超重力处理引起绿豆同工酶结构和活性的改变。     

UV-B辐射对拟南芥POD、SOD同工酶的影响

选择哥伦比亚生态型(Columbia-0)的野生型拟南芥(Arabidopsis thaliana L.)为供试材料,研究了其POD、SOD同工酶对不同剂量UV-B胁迫的响应.结果显示,POD和SOD同工酶活性与UV-B辐射的剂量关系密切.POD、SOD同工酶活性在低剂量UV-B辐射时呈增加趋势,在中、高剂量UV-B辐射时活性则呈下降趋势.在UV-B胁迫下,POD同工酶酶带没有发生变化;SOD同工酶酶带在胁迫下,出现了新的酶带(SOD4、SOD5).说明拟南芥在受到低剂量的UV-B胁迫下,可以通过提高自身的保护系统来抵御外界不良环境的影响;而受到较高剂量的UV-B胁迫时,会破坏植物的保护系统,造成不可逆的损伤.     

烟草感染青枯菌前后POD及同工酶的变化

【研究目的】对烟草感染青枯菌前后的根、茎、叶内过氧化物酶(POD)及同工酶进行比较研究,为抗病育种提供理论基础。【方法】采用比色法测POD酶活,用聚丙烯酰胺凝胶电泳(PAGE)进行POD同工酶的比较。【结果】对烟草感染青枯菌前后,酶活性测定结果表明,烟草叶部POD活性显著高于根部和茎部。烟草接菌后,根、茎、叶内的POD活性均在第3天升至最高,而叶内比对照增高了29.67%。同工酶电泳分析表明,烟草接菌后的同工酶谱带,在条带的数目、种类和各条的相对活性上表现出差异。【结论】POD作为植物的保护酶类,随植物抵抗疾病的过程变化而变化,故与植物抗病性密切相关。     

zyxin基因SNP(exon 1)与京海黄鸡生长和屠体性状的相关性

以京海黄鸡为试验材料,采用PCR-SSCP技术检测斑联蛋白基因(zyxin)8个外显子SNP(单核苷酸多态性),探讨zyxin基因的多态性与鸡生长和屠体性状之间的关系。结果发现zyxin基因外显子1有1个SNP突变位点,表现为3种基因型,分别用AA、AB和BB表示。统计分析结果表明:BB型的公鸡腹部脂肪重、屠宰率显著高于AA型与AB型(P<0.05),AA型与AB型差异不显著。AB型的母鸡心重、胸肌率、腿肌率显著高于AA、BB型,而半净膛率、全净膛率低于AA、BB型。公鸡、母鸡的生长和屠体性状在不同基因型间比较,除了半净膛率以外,其余各指标均存在显著或极显著差异。以上研究结果表明,zyxin基因可能是影响京海黄鸡部分内脏组织、器官和屠宰率等相关指标的基因。     

猪MYOG基因遗传多态性与胴体及肉质性状的相关性

通过测定约180日龄的8个群体(梅山、大白、长白、大白×梅山F1代、梅山×大白F1代、长白×大白F1代、大白×长白F1代以及大白×梅山F2代)459头猪胴体性状、肉质性状以及肌细胞生成素基因(MYOG)多态性,分析MYOG在猪群体中的遗传效应。结果表明：MYOG在猪群体中呈现3种基因型,即AA、AB、BB;群体瘦肉率的提高与BB基因型显著相关,相较于AA与AB基因型,其提高幅度分别为3.57%和3.28%;在大白×梅山F2代群体中,MYOG的基因型与背最长肌pH值、股二头肌pH值呈显著相关;在大白×梅山F1代群体中,MYOG基因型与股二头肌大理石纹MM2呈显著相关。     

多酚氧化酶在小麦植株体内的分布

分析小麦灌浆后期植株籽粒、茎秆、叶片、根多酚氧化酶(PPO)活性及同工酶的差异,结果表明:小麦植株自上而下,PPO活性和同工酶种类有下降的趋势,即剑叶＞籽粒＞穗颈＞第2节＞第3节＞第4节＞根,同工酶种类以籽粒中最为丰富,共有9条谱带.     

Polymorphism Analysis on Partial Sequence of Pig Obese Gene of Different Breeds by PCR-SSCP

Obese gene (ob gene) was identified in mouse adi- pose tissues in 1954, and it has an autosomal reces- sive genetic pattern[1]. Mouse ob gene is located on chromosome 6, whose mutation will lead to seriously genetic fatness. Human ob gene is mapped on q31…     

不同除草剂对不同谷子品种POD同工酶的影响

为了解决生产上谷子杂草多的困扰,也为谷子种植上选用合适的除草剂提供理论依据,试验采用完全随机设计,以4个谷子品种和2种除草剂为研究材料,通过扑草净土壤施药、扑草净茎叶施药、莠去津土壤施药、莠去津茎叶施药和清水喷施,在谷子苗期、拔节期、孕穗期对POD同工酶活性测定并分别进行比较。结果表明,不同除草剂处理下,苗期谷子POD活性变化趋势基本一致,扑草净土壤处理莠去津土壤处理莠去津茎叶处理对照扑草净茎叶处理。拔节期和孕穗期的POD活性之间差异都不显著,说明谷子生长达到了正常水平。这与生理指标表现出来的规律基本相似。研究证明,4个品种谷子都能适应莠去津、扑草净2种除草剂胁迫而产生相应变化,从而调节细胞代谢以抵抗这种胁迫。     

Kinetics of acetylcholine synthesis and hydrolysis in myasthenia gravis

Kinetic studies for choline-O-acetyltransferase (CAT) (E.C. 2.3.1.6) and acetylcholinesterase (E.C. 3.1.1.7) were performed on serum, skin fibroblasts in culture, and biopsied sartorius muscle from normal and myasthenic subjects. There was a significant decrease of CAT activity per milligram of protein in myasthenic muscle compared to normal muscle, and there was no difference in acetylcholinesterase activity per milligram of protein in the same muscle homogenates. Substrate concentration curves for acetyl coenzyme A and CAT also showed a significant reduction in the maximum rate of product formation (V(max)) per milligram of protein between myasthenic and normal muscle. It is postulated that binding of substrate to CAT is being inhibited by an inhibitor present in muscle.     

Peafowl lactate dehydrogenase: problem of isoenzyme identification

Peafowl, like other vertebrates, contain multiple forms of lactate dehydrogenase. The electrophoretic properties of the peafowl isoenzymes are unusual in that the isoenzyme from heart tissue can be either more or less anodic than that of muscle, depending on the pH. This finding focuses attention on the problem of isoenzyme identification. It is suggested that isoenzymes be identified on the basis of properties that are chemically and biologically more significant than electrophoretic mobility.