高压处理对肌球蛋白质的影响

从牛骨骼肌中提取肌球蛋白质后进行纯化,并用不同程度的压力(100,200,300,400 MPa)进行处理,再与未处理样品进行比较。采用SDS—PAGE分析了解压力对肌球蛋白的影响。从SDS—PAGE可观察到,随着压力的升高,肌球蛋白质条带逐渐变淡,压力为400 MPa时,13.8 kDa的肌球蛋白轻链几乎已经消失,而200 kDa的肌球蛋白重链下方出现了一条比较模糊的其他蛋白质条带。     

大豆质核互作雄性不育系W931A及其保持系的差异蛋白质组学比较分析

采用SDS—PAGE与LC—MS／MS联用方法．对大豆质核互作雄性不育系W931A及其同型保持系W931B不同器官的蛋白质组进行比较性研究．结果发现不育系W931A与其保持系W931B种子和苗期叶片的蛋白质SDS—PAGE电泳图谱基本没有差异带．二胞花粉期花药SDS-PAGE图谱间有3条差异带。雄性不育基因表达具有时空和器官特异性,与育性有关的蛋白主要在花药中表达。对差异表达的蛋白如ADP-葡萄糖焦磷酸化酶、NBS—LRR疾病抵抗蛋白质的同源物、ATP合成酶b亚基、硫氧还蛋白过氧化物酶、类锌脂蛋白等进行功能分析．推测不育系W931A的雄性不育可能与淀粉合成受抑制、能量代谢紊乱和细胞凋亡有关。     

小麦高代重组自交系中HMW-GS基因的分子鉴定

以JY970012群体内11个F8代重组自交系为材料，采用SDS—PAGE技术和AS—PCR技术对HMW—GS组成进行鉴定。结果表明：每个重组自交系在Glu-A1、Glu—B1、Glu—D1位点上HMW—GS组成稳定。F8代，AS—PCR技术在苗期鉴定表明：含有1亚基的株系具有约1500bp的特异带，而含有null亚基的株系没有此带；含有null亚基的株系在约920bp处出现特异带，而1亚基没有。SDS—PAGE和AS—PCR两种技术鉴定结果一致，同时，PCR技术简便、快速、准确、科学，可检测优质亚基基因，为小麦品质选育提供理论依据。     

棉花黄萎菌落叶型菌系毒素纯化及抗血清制备

用SephacrylS 200HR层析柱纯化了V991菌系毒素,经叶片针刺涂抹方法确认其致萎功能后,免疫大白兔制备了抗血清。用间接ELISA方法对强、中、弱3种不同致病力共10个菌株的培养液进行了特异性检测,结果表明:抗血清具备一定生理型特异性,能够准确检出全部强致病力落叶型菌系,可作为不同生理型划分的指标之一。用毛细管电泳、SDS PAGE和Western杂交方法检测了V991毒素的组分:在中性条件下,毒素在毛细管电泳时存在2个峰;SDS PAGE将毒素分为8条带,其中4条主带具有强烈的致萎性;Western点杂交的反应强度与相应蛋白回收带的致萎功能之间明显相关。     

未稳定和稳定处理的米糠蛋白质浓缩物的制备及其特性

通过碱提取和等电沉淀从未稳定的和加热稳定处理的米糠中提取蛋白质浓缩物，并对它们的特性进行了比较。经稳定处理的米糠在各种粒度下（75～150μm）蛋白质提取性较低。在PH8．0时，未稳定的米糠蛋白质浓缩物（URBPC）和稳定的米糠蛋白质浓缩物（SRBPC）对氮的溶解性最大。URBPC（71．5％蛋白质）和SRBPC（50．9％蛋白质）表明氨基酸含量存在差异。两种蛋白质浓缩物SDS－PAGE表明有一些共同成分，但SRBPC中缺少某些成分。可以肯定由于米糠经加热稳定，蛋白质变性，从而降低了蛋白质的提取，并影响了蛋白质的品质。未稳定和稳…     

营养对五龙鹅基因表达影响的研究

对著名的地方特色品种五龙鹅进行不同营养水平试验,用聚丙烯酰胺凝胶电泳(SDS)PAGE技术,分析血液中脂酶(EST)、乳酸脱氢酶(LDH)及蛋白质的多态.结果表明 试验组蛋白质结构新出现S1,S3,S4亚基组分、存在丰富的多态变化.EST同工酶快区Es-1位点,由于等位基因控制,消减Es-1D、Es-1F同工酶谱带,其活性下降20.7%.LDH同工酶基因表达调控A和B亚基的构成,出现LDH1亚基,其活性上升99.6%.提示营养环境直接或间接调控基因表达水平,与生长性能呈正相关.     

利用SDS-PAGE鉴定水稻品种(组合)的真实性和纯度

利用本实验室完善后的SDS—PAGE，分析了8个水稻品种（组合）的贮藏蛋白，结果表明，品种（组合）间贮藏蛋白带谱表现出明显的多态性；并在实际检验工作中，运用水稻品种（组合）贮藏蛋白SDS-PAGE带谱的多态性，进行了种子的真实性或纯度鉴定。     

棉苗抗冷性与低温诱导蛋白关系的研究

以具有显著抗冷性差异的早熟和晚熟两种陆地棉品种为材料,以昼夜不同形式的低温处理其幼苗后,测定其可溶性蛋白含量;并采用SDS聚丙烯酰胺凝胶电泳法(SDSPAGE)对蛋白质进行分离,发现抗冷性强的新S29品种的幼苗经适当的低温处理后有一条分子量约123kd的低温诱导蛋白(即抗冻蛋白)谱带产生,而抗冷性弱的晋棉6号则没有。较详细地分析了不同温度处理对棉苗中可溶性蛋白质含量和种类的变化情况及其与品种抗冷性间的关系,并从理论上探讨了与其相关的可能机理。     

用可溶性蛋白电泳法鉴定玉米小斑病菌生理小种

分别从玉米小斑病菌T、C、O三个生理小种的菌丝中提取可溶性蛋白,进行聚丙烯酰胺凝胶电泳(PAGE)和SDS聚丙烯酰胺凝胶电泳(SDS-PAGE),经多次比较试验,T、C、O小种的蛋白质图谱主带基本相同,个别带存在一定差异,而同一生理小种不同菌株间差异甚小.建议运用病原菌蛋白电泳的方法,作为小种分类、病害预测的手段之一.     

水分亏缺条件下豌豆肌动蛋白异型体PEAcII基因的表达

以培养3～4 d的豌豆幼苗为试验材料,以18S rRNA基因为内参基因,豌豆幼肌动蛋白异型体PEAcII基因为检测基因,在PEG6000模拟水分亏缺条件下,采用半定量逆转录聚合酶链式反应(RT-PCR)体系检测肌动蛋白异型体PEAcII基因在豌豆幼苗中的表达情况。在正常供水条件下,PEAcII在处理的第8 h表达量最高,水分胁迫下,PEA-cII基因在处理的0.5 h表达量下降,在处理的1 h时表达量最高,而后下降。无论是正常供水还是水分胁迫条件下,PEAcII基因的表达都是一个动态变化的过程。试验结果表明,肌动蛋白异型体PEAcII表达量的高低可能与豌豆幼苗适应水分亏缺的生理过程有关,这对于研究豌豆幼苗抗旱性机理以及检测豌豆幼苗的抗旱性强弱具有重要意义。     

聚丙烯酰胺凝胶电泳鉴定烟草品种

在本实验条件下，得到的8个烟草品种种子的可溶性蛋白质的SOD-聚丙烯酚胺凝胺电泳（SDS-PAGE）和聚丙烯酚胺等电聚焦电泳（IEF-PAGE）图谱，分辩率高，重复性强，其多态性条带分别占61.5%和52.6%,各品种的电泳图谱可以相互区别,表明种子的可溶性蛋白的SDS-PAGE和IFF-PAGE图谱可作为烟草品种鉴定的蛋白质指纹图谱.     

Allelic variants of granule-bound starch synthase proteins in European bread wheat varieties

The composition of 324 European wheat cultivars were analysed at the three granule‐bound starch synthase (GBSS I) loci. Protein separation was first made by sodium dodecyl sulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE. A specific two‐dimensional (2D) electrophoresis (immobilized pH gradient × SDS‐PAGE) using an Immobiline dry strip in the first dimension was developed to resolve the GBSS I proteins more clearly and to confirm some results. Very low polymorphism was found. Among the 324 cultivars analysed, only one carried a Wx‐A1 null allele (Wx‐A1b) and none was found to have the Wx‐2D null allele. As described in the literature the Wx‐B1 locus was more polymorphic and the null allele was encountered in 11 cultivars. The use of 2D electrophoresis allowed us to find another type of variant which presented as having thicker band with same mobility as the Wx‐D1 protein in SDS‐PAGE. Twelve per cent of the cultivars analysed presented this band and could have been previously mistaken for cultivars carrying the Wx‐B1 null allele. Indeed this band probably corresponded to the Wx‐B1? or Wx‐B1e allele overlapping with the Wx‐D1a allele in SDS‐PAGE.     

小麦谷蛋白聚合体的MS-SDS-PAGE及其与面包烘烤品质的关系

选用品质(微量SDS沉淀值、稳定时间)优劣不同的10个"9311168///81831-1/pernell//京411"的F6品系, 探索多层浓缩胶SDS-PAGE(MS-SDS-PAGE)的实验方法, 初步探讨可溶性谷蛋白的分子量分布状况和不溶性谷蛋白(GMP)与面包烘烤品质的关系. 结果表明, 通过SDS-磷酸缓冲液可将谷蛋白聚合体分成两部分: 分子量较小的SDS-可溶性谷蛋白     

小麦幼胚无性系麦谷蛋白亚基变异研究

对通过辐射诱获得的323个无性系进行了SDS－PAGE电泳分析，结果表明，小麦储藏蛋白变异几率随愈伤辐射剂量的增强而增加，但是随辐射剂量的增加，愈伤存活率，绿苗再生率了随这剧烈降低。同时分析了麦谷蛋白变异类型及其与植株形态变异的关系，提出了小麦幼胚愈伤组织辐射诱变的适宜剂量。     

Studies on the Mechanical Properties of Erythrocyte membrane for Hereditary Spherocytosis

A SDS PAGE technique is adopted to investigate the change of membrane cytoskeleton protein for hereditary spherocytosis(HS).The membrane vicoelastic properties are determined by the micropipette aspiration technique.The results show that the membrane elastic modulus of HS erythrocyte hase no remarkable difference but the viscosity is significantly higher than normal cells and the deformability of HS erythrocyte is reduced.It is discovered that spectrin,band 3 and band 5 of HS erythrocyte are partly deficient;the membrane thiol groups concentration of HS erythrocyte is lower than normal cells.These results suggest that the integrity of the struture and ingredient of the cytoskeleton protein networks plays an important role in deformability of erythrocvte.     

The effect of indirect tests for grain quality on the grain yield and industrial quality of bread wheat

The objective of this study was to examine the effect of selection using three indirect tests for grain quality on grain yield and dough, and baking properties, measured as alveograph strength, alveograph tenacity/extensibility ratio and loaf volume. The three tests were flour protein content, flour sedimentation and high molecular weight glutenin subunits. Of the indirect tests used for grain quality, sodium dodecyl sulphate (SDS)‐sedimentation allowed the highest intensity of selection for a combined trait index of the target grain quality and grain yield characteristics. The top 48% of the material could be retained on the basis of SDS‐sedimentation, resulting in retention of atleast two‐thirds of the top 10% of genotypes for the combined trait index. Flour protein percentage, a weighted high molecular weight glutenin index and an index combining all the indirect tests—flour protein, SDS‐sedimentation and high molecular weight glutenins—gave selection intensities of 61%, 64% and 55%, respectively, for the combined trait index. If the objective of selection is dough strength alone, then a weighted index of all indirect traits (flour protein, SDS‐sedimentation and high molecular weight glutenins) provided the highest selection intensity (26%). Other selection intensities for individual target traits were 24% for the prediction of loaf volume from flour protein, 40% for the prediction of tenacity/ extensibility ratio using SDS‐sedimentation and 68% for the prediction of grain yield using SDS‐sedimentation.     

小麦高分子量谷蛋白亚基与小麦品质性状关系的研究

采用SDS-PAGE方法, 通过对4个优质亲本与3个农艺亲本间杂交获得的F2群体的每一单株及其亲本进行小麦高分子量谷蛋白亚基(HMW-GS)组成分析, 并对每一F2单株上F3籽粒群体的高分子量谷蛋白亚基组成与其籽粒蛋白质含量、 SDS-沉降值的关系进行研究,结果表明：小麦高分子量谷蛋白亚基组成不同的群体间籽粒蛋白质含量差异不显著     

Variation in seed proteins from mutagen-treated cultivars and selected lines of Vigna unguiculata L. Walp

The storage protein profiles of the seeds of two IITA cowpea cultivars (‘IT84E–124’ and ‘Vita 7’) exposed to three mutagens—sodium azide (NaN₃), ethyl methane sulphonate (EMS) and ⁶⁰Co gamma rays—and those of 18 selected M₃ lines were investigated by sodium dodecyl sulphate (SDS)-polyacrylamide gel electrophoresis (PAGE). The total seed protein, globulin and albumin fractions showed differences in number and intensity of subunit bands, but the least differences were found in the albumin fraction. Both high and low molecular weight bands were observed, the highest being 94.5 kDa and the lowest 12.0 kDa.‘Vita T showed less variability compared with ‘IT84E–124’, as indicated by the relative similarity indices (S.I.) of the two cultivars. The lowest S.I. of 0.200 was found among ‘IT84E–124’ lines while the lowest S.I. among ‘Vita T lines was 0.4706. A number of lines with particular traits were found to be characterized by the presence of specific polypeptide bands. This study demonstrates that induced mutation could create additional variability to supplement existing germplasm and that SDS-PAGE is a useful tool for discriminating and estimating genetic similarities among selections.     

利用种子蛋白聚丙烯酰胺凝胶电泳鉴定向日葵种子纯度的研究

采用种子贮存蛋白SDS-PAGE技术，对13份向日葵杂交种和自交系进行了鉴别，结果表明，它们的种子蛋白电泳图谱差异明显，特征谱带稳定。种子蛋白电泳鉴定与田间小区种植鉴定种子纯度结果之间存在极显著的线性正相关性。该技术可作为向日葵杂交种和自交系种子纯度鉴定的一种新手段。