聚丙烯酰胺凝胶电泳的一种辅助辨型方法

聚丙烯酰胺凝胶电泳（PAGE）是实验室常用的基本鉴定方法之一。但由于做胶方式以及各实验室仪器设备良莠不齐，经过一系列纷繁复杂的操作，电泳结果未必尽如人意。其中凝胶带型的判断就是一个较为困惑的问题，一般的聚丙烯酰胺胶跑出的条带总体上都有弧度，越是靠近凝胶两侧越是严重。在本实验中，我们设计了一种较为简单易行的方案，即将难以区分的条带样品做成混合样marker来辅助辨型，通过调整电压和凝胶浓度来达到区分条带的目的。我们用该方案可以将相差0～3bp大小的条带区分开来。     

Changes in meat texture of three varieties of squid in the early stage of cold storage

ABSTRACT: The present study examined the changes in texture and protein components during cold storage of different squid varieties. Raw oval squid, Japanese common squid and arrow squid were sliced fresh and the muscles were stored at 4°C for 0, 4, 8, 12, 18, 24, 48 and 120 h. The rheological measurements, protein components and amounts of collagen were examined. The adhesiveness of each squid increased significantly in the early stage of cold storage. In all varieties, penetration decreased at 4 h, which is considered to be rigor mortis, then increased. The amounts of total collagen, 20°C water-soluble collagen and 70°C water-soluble collagen did not change significantly in each variety during cold storage. Sodium dodecylsulfate–polyacrylamide gel electrophoresis (SDS-PAGE) pattern showed that the 580 kDa component gradually disappeared up to 48 h. The correlations between the amounts of 580 kDa component and adhesiveness or firmness were high. Models of fit based on chemical kinetics accurately expressed the behavior of adhesiveness, firmness and penetration showing that 63.2% of adhesiveness changes occurred in 13–19 h and that 63.2% of firmness changes occurred in 18–24 h.     

应用核酸斑点杂交法检测鹅细小病毒(GPV)

利用鹅细小病毒 (GPV)核酸探针 ,设计并建立了核酸斑点杂交法 ,用以检测 GPV核酸。该方法特异性与敏感性均较好 ,其敏感度可达 3pg。应用该方法对 4例病变典型的小鹅瘟患鹅组织器官进行检测 ,结果表明 ,患鹅脏器中 ,小肠、心、肝、胰、脾组织含毒量较高 ,其中小肠组织含毒量最高 ,而脑组织中含毒量较低。应用琼脂扩散试验验证了核酸斑点杂交法的检测结果。     

家蚕血液酯酶同功酶的研究

采用不连续聚丙烯酰胺凝胶电泳法.测定了24个家蚕品种及杂交种从三龄到后蛹不同发育时期的血液酯酶同功酶酶谱.结果表明:家蚕血液中存在着丰富的酯酶同功酶酶带类型,整个试验可观察到22条酶带,根据其迁移率及染色特点可分为A、B、C、D、E五大染色区.在不同发育阶段中共有13条酶带经常显现,后蛹期主要同功酶酶带活性降低,但非主带A-1区酶带却清楚地显示出来.在幼虫阶段,酶带的数目及活性防着食下量的增多而增人.并观察到A-2-b带与B-2带的表现最为突出,推测A-2-b带可能是家蚕血液部酶同功酶的基本酶带.进一步分析表明,A-2-b带活性与大多数经济性状间呈高度的正相关性.     

荞麦种子蛋白质组分分析

研究了不同品种荞麦种子中蛋白质组分情况，结果表明：养麦种子蛋白质主要由清蛋白、球蛋白、谷蛋白和醇溶蛋白构成。而不同类型蛋白质在总蛋白质中所占比例有明显差异，清蛋白是荞麦种子中的主要蛋白质，占总蛋白质含量的71．4％左右，其中以平荞最高。醇溶蛋白在荞麦种子总蛋白中所占比例较小，平均约为7．4％，榆养2号最高。球蛋白和谷蛋白所占比例非常接近，其中球蛋白平均约占9．1％，最高为日本大粒养。谷蛋白大约占总蛋白的12．1％，以京养1号最高。     

连作对棉田土壤枯、黄萎病菌数量及细菌群落的影响

采用选择培养法测定新疆库尔勒不同连作年限棉田土壤枯、黄萎病病菌数量的变化,运用PCR-DGGE(Polymerase chain reaction-denatured gradient gel electrophoresis)技术分析棉花连作对土壤细菌群落结构和多样性的影响。结果表明,不同连作年限棉田的枯萎病菌和黄萎病菌量具有相似的变化特征,即在5~20年连作年限内,土壤中病原菌量随着连作年限的延长而增加,20年连作土壤病原菌数量达到最高峰;连作超过20年,土壤菌源数量开始下降。不同连作年限棉田土壤中枯、黄萎病病原菌的数量与发病株率呈显著正相关关系。连作棉田土壤细菌群落的DGGE图谱分析显示,连作棉田土壤细菌种群的组成、结构、丰富度均发生变化。连作5~40年的棉田土壤细菌种群Shannon-Wiener多样性指数(H')、均匀度(EH′)和丰富度(S)指数均降低,且连作年限越长降低越突出。     

Inheritance of peroxidase isozymes in mulberry (Morus spp.)

Summary Electrophoretic variants of peroxidase in mulberry (Morus spp.) were demonstrated by thin-layer gel isoelectric focusing. Of these variants, three isozyme band groups were found to be controlled by codominant alleles at a single locus. The gene symbol Px ₁ was given to this locus, with alleles Px ₁ ¹ and Px ₁ ² assigned to the A6-A7-A8 and A7-A8-A9 band groups, respectively. The A6-A7-A8-A9 band group proved to be controlled by the Px ₁ ¹ and Px ₁ ² heterozygote.Additional experiments showed that among the three banding types, there were no statistically significant differences in leaf blade length, leaf blade width, length-width ratio of leaf blade, internode length, phyllotaxis, leaf shape, tree vigor and resistance to powdery mildew, but there were significant differences in leafstalk length.     

盐胁迫对燕麦种子萌发期蛋白质组影响的研究

为探析盐胁迫下燕麦种子萌发时蛋白质的差异表达及其相关生理功能分析,比较了燕麦在不同浓度氯化钠溶液处理下种子的萌发及幼苗生长能力,然后采用二维凝胶电泳结合基质辅助激光解析飞行时间质谱的蛋白质组学方法分析种子在0和1%氯化钠溶液处理种子萌发时的蛋白质组差异表达。结果表明:随着盐胁迫的加剧,燕麦种子的萌发力和成苗率降低。蛋白质组分析显示:1%氯化钠溶液处理使得燕麦种子蛋白质组二维凝胶中11个蛋白质点表达发生变化,其中10个蛋白质点表达量上调。质谱鉴定得到3个可靠蛋白,分别为:燕麦蛋白、燕麦蛋白N9和蛋白质二硫键异构酶。这些蛋白的差异表达可能与盐分胁迫下燕麦种子萌发的生理活动有关联。     

适用于水稻微粒体蛋白的双向电泳技术及苯达松抗性相关蛋白的初步分析

针对不同来源的样品，建立相应的双向电泳(two-dimensional gel electrophoresis, 2-DE)技术是开展蛋白质组学研究的基础。以野生型水稻(O. sativa L.)农林8号(N8)及其苯达松敏感致死(bentazon sensitive lethal, bsl)突变体农林8号m (N8m)为材料，通过优化条件，建立了适合于水稻微粒体蛋白的双向电泳技术。并通过双向电泳图谱的比对，共找到15个差异蛋白点，分为供试材料间差异、苯达松处理前后差异和N8在苯达松处理后特异表达等3种类型。结合苯达松抗性比较及苯达松残留分析，推测材料间差异是因γ射线辐照引起基因结构变异，从而影响了基因的表达；苯达松处理前后差异与氧自由基清除酶系或系统获得抗性相关酶系有关；N8在苯达松处理后特异表达与苯达松代谢有关。