冷刺激对两种泥鳅血液蛋白质的影响

采用SDS-聚丙稀酰胺凝胶电泳技术,探讨冷刺激对泥鳅血液蛋白质的影响.结果表明,泥鳅和大鳞副泥鳅的血液蛋白质发生了变化,即产生了冷激蛋白;同种泥鳅,控制产生冷激蛋白的特定基因在一定的刺激强度和刺激程度下,产生冷激蛋白的时间和种类不同;不同种泥鳅,在相同低温处理条件下,产生冷激蛋白的时间和种类也不一样.本研究不仅为研究冷激蛋白提供了前提,而且为两种泥鳅的人工养殖提供了参考.     

恩施碎米荠种子蛋白质双向电泳初探

对湖北恩施高硒地区生长的具有超富硒能力的植物恩施碎米荠的种子蛋白质用TCA-丙酮沉淀提取,等电聚焦采用固相梯度胶条,双向电泳分离蛋白质.结果表明,蛋白质样品制备的质量高;银染胶图背景干净,蛋白点清晰,且呈现较好的聚焦结果,无明显拖尾或横纹现象;蛋白质集中分布在pH5～10的范围之内,分子量主要在20～60 kμ的范围之内.     

牙鲆血清免疫球蛋白的分离纯化及部分特性分析

运用Sephacryl S-200凝胶层析和HiTrap rProtein ASepharose亲和层析2种方法对牙鲆(Paralichthys olivaceus)血清免疫球蛋白进行分离纯化,结果表明,牙鲆免疫球蛋白分布于33%~50%的硫酸铵饱和溶液中,其中45%的分离效果最好。凝胶层析和亲和层析样品均出现2个蛋白峰,用还原SDS-PAGE检测确定牙鲆免疫球蛋白存在于第2个蛋白峰中。牙鲆免疫球蛋白重链分子量约为75.4 kD,轻链分子量约为29.9 kD和28.2 kD,推测牙鲆血清免疫球蛋白的分子量为836 kD。制备了兔抗牙鲆免疫球蛋白多克隆抗体,免疫双扩散法检测多克隆抗体效价为1∶32,免疫斑点法检测多克隆抗体效价至少为1∶1 600。运用免疫印迹法(Western-bloting)检测了兔抗牙鲆免疫球蛋白多克隆抗体的特异性,实验证明该抗体与牙鲆全血清中免疫球蛋白重链、轻链反应均成阳性。     

单细胞凝胶电泳检测除虫脲光解产物对小鼠肝脏DNA损伤研究

采用单细胞凝胶电泳法对不同浓度除虫脲经紫外光照射后的混合物可能导致小鼠肝脏细胞DNA损伤的作用进行了研究,应用CASP软件分析彗星图像,得出彗星尾部DNA百分含量、彗星尾长、尾距和Olive尾距4个指标.通过SPSS软件分析表明,其光降解混合物导致的彗星拖尾现象与空白组有显著区别,且在一定浓度范围呈现剂量-效应关系,表明不同浓度除虫脲经光降解后其混合物具有一定的致突、致癌性,且毒性随着浓度的降低而减小.     

水稻穗上发芽的差异蛋白质分析

采用IEF-SDS-PAGE双向电泳技术,对穗上发芽的水稻胚芽进行蛋白质分离,获得清晰的电泳图谱。结果表明:同一品种未发芽和穗上发芽的水稻胚芽蛋白质电泳图谱基本一致,体现了其蛋白质组分的稳定性,同时未发芽和穗上发芽的水稻胚芽蛋白质也具有各自的特异性。从图谱中得到50个差异表达的蛋白质,其等电点多集中在pH5.0-6.5之间,分子量多集中在14.4-67.0ku之间。这些差异蛋白的分析结果为今后进一步研究水稻穗上发芽的分子机理奠定了试验基础。     

黄土丘陵区垄沟改良措施对玉米水分利用效率的影响

选择延安地区作为黄土丘陵区的代表,在延安地区应用地膜、液态地膜、保水剂、PAM(土壤改良剂)和间作套种措施优化大垄沟种植方式种植玉米,分析了不同优化措施种植玉米的水分利用效率。结果表明,地膜、保水剂、PAM和间作方式有利于大垄沟较好地保持土壤水分,其保水能力分别比平种高出17.4%、8.5%、11.6%、5.3%;液态地膜、地膜、间作和PAM表现出很好的增产效果,其产量分别比平种高出77.7%、72.0%、64.1%、30.8%。     

甘草愈伤组织蛋白质组双向电泳技术的建立

以来源于野生乌拉尔甘草的愈伤组织为试材,对甘草愈伤组织蛋白质的提取、SDS-PAGE电泳、上样量、染色方法等关键步骤进行了优化,结果表明:采用改良丙酮法可提高提取液中蛋白质的含量及单向电泳图谱的分辨率;17 cm IPG胶条上样量为450μg时,采用银染法提高了双向电泳图谱的分辨率及分离效果,获得了1 132个蛋白点。建立了一套适用于乌拉尔甘草的愈伤组织蛋白质组分析的双向电泳方法。     

茭白黑粉菌脉冲电泳分析方法

采用脉冲电泳法对茭白黑粉菌染色体进行核型分析。分析结果表明,茭白黑粉菌脉冲电泳样品处理方法包括原生质体的制备、电泳参数的设置等,对试验结果有一定影响,在制备电泳样品时,包埋的原生质体或孢子的浓度不能太低,蛋白酶K的浓度不能过高,低熔点琼脂糖的浓度不能太低,此外还应确保电泳槽水平,并且电泳结束后及时取出凝胶。     

番茄青枯病抗性两个相关基因功能的初步鉴定

对抗性番茄材料‘Hawaii 7996’分别接种强致病力青枯菌（RsH）和中等致病力青枯菌（RsM）后,利用双向电泳技术找到了两个差异表达蛋白质--谷胱甘肽S转移酶L3和Remorin 1蛋白。在利用携带八氢番茄红素脱氢酶基因PDS的TRV重组病毒载体通过叶片浸润法证明VIGS体系可行后,进一步利用该技术沉默上述两个蛋白质的对应基因GST-L3和Rem-1,并用半定量RT-PCR检测沉默效果。结果表明,分别沉默GST-L3和Rem-1的番茄植株抗性减弱,在接种中等致病力青枯菌（RsM）后青枯病发病率和病情指数都明显高于对照植株,说明基因GST-L3和Rem-1与番茄对青枯病的抗性相关。     

Distribution and Genotypic Characterization of Campylobacter jejuni Isolated from Poultry in Split and Dalmatia County,Croatia

Consumption of poultry contaminated with Campylobacter jejuni has been recognized worldwide as the leading cause of campylobacteriosis. Therefore, the aim of our study was to investigate the prevalence and genotype diversity of Campylobacter jejuni in poultry meat intended for consumption in Split and Dalmatia County, which is the second biggest County in Croatia. Furthermore, we also wanted to discover possibly stable clones of C. jejuni appearing in different samples and periods of time, which would indicate their ability to persist in or adapt to poultry. In the period from March 2008 until June 2010, 834 samples of poultry from various sources were examined using a surface swab technique. Isolation of C. jejuni was performed by Preston broth and Karmali agar. Identification of the isolates was carried out using biochemical tests. C. jejuni was found in 84 of 574 chicken samples (14.6%) and in nine of 260 samples of turkey (3.5%). Pulse‐field gel electrophoresis (PFGE) was used to analyse 61 obtained isolates using SmaI and KpnI. Of 22 different macrorestriction profiles (MRP) that were found, five were detected in poultry from both different locations and periods of time. Samples from 11 locations were found to be contaminated with more than two different genotypes of C. jejuni. Interestingly, the same MRP were found both in poultry declared to be of domestic origin and in the poultry imported from abroad. The prevalence of C. jejuni in poultry samples was in accordance with previously reported results. Genotypic analysis indicated that the population of C. jejuni in Split and Dalmatia County was diverse and that multiple strains of C. jejuni could be found in the same poultry samples. Furthermore, the same genotypes were identified from the samples obtained from different locations and periods of time, which could support the theory of a global existence of certain MRP that are able to persist in or adapt to poultry.