含FMDV 2A序列PRRSV GP5和M蛋白重组腺病毒的构建与鉴定

【目的】构建表达PRRSV GP5和M蛋白、且二者通过口蹄疫病毒(FMDV)2A序列连接的重组腺病毒,以期实现2个蛋白的同时表达,发挥GP5蛋白的病毒中和优势和M蛋白的细胞免疫优势作用。【方法】利用RT-PCR、间接免疫荧光和Western blot等方法,对获得的重组腺病毒(rAd-GP5-2A-M)进行检测。【结果】检测结果均证明,GP5-2A-M蛋白不仅获得了正确表达,而且能自我裂解为GP5和M蛋白。【结论】在GP5和M蛋白之间插入具有自我裂解功能的FMDV 2A策略是切实可行的,并为PRRSV多个蛋白的共表达奠定了基础,也为其他病毒基因工程疫苗的研制提供了全新思路。     

Antimicrobial activity of the essential oil of Centaurea aladagensis

Composition of the water-distilled oil of the aerial parts of Centaurea aladagensis, endemic in Turkey, was analysed by GC-MS. Hexadecanoic acid (39.3%), caryophyllene oxide (6.6%) and hexahydrofarnesyl acetone (4.3%) were found as main constituents in the oil. The oil was tested against 7 human pathogenic microorganisms.     

辐照对冷鲜熟制水饺微生物安全质量的影响

本试验主要研究了辐照对冷鲜熟制水饺中可能存在的致病微生物肠炎沙门氏菌、金黄色葡萄球菌和李斯特菌存活状况的影响。研究发现,肠炎沙门氏菌、金黄色葡萄球菌、李斯特菌的D10值分别为0.31、0.44和0.45kGy。随着辐照剂量的增大和贮存期的延长,不同致病菌数量变化趋势存在差异。4kGy辐照可保障冷鲜熟制水饺中3种致病菌达到不得检出的要求。     

禽大肠杆菌O2、O78及其耐药、融合菌株的外膜蛋白免疫研究

为了研究大肠杆菌O2、O78及其耐药、融合菌株外膜蛋白的免疫原性,用Sarkosyl处理和超速离心技术相结合的方法分别提取禽大肠杆菌强毒株O2、O78,耐药弱毒株O2(Nor^rChl^s)、O78(Not^sChl^r)及其融合双价弱毒株O2.78(Nor^rCll^r)等5个菌株的外膜蛋白(OMPs),经SDS-PAGE,结果均出现1条相同的相对分子质量在31000-43000之间的主要OMPs条带,表明5个菌株存在相同的OMPs成分,将3个耐药菌株的OMPs分别免疫小白鼠和鸡,结果均产生对O2、O78良好的同源及交叉免疫保护效果,用各耐药菌株的OMPs做包被抗原,ELISA方法都能交叉检测3个耐药菌株OMPs免疫鸡血清中IgG水平,且各IgG的消长曲线基本一致。     

Pathogenicity of Bipolaris sorokiniana isolates from wheat roots, leaves and grains in Mexico

Pathogenicity of 27 Bipolaris sorokiniana isolates from roots, leaves and grains of spring wheat, collected at a site in Mexico, showed no clear differences between groups of isolates, based on lesion density after seedling inoculation of the susceptible cultivar Ciano T-79 under controlled conditions. Results were variable when tests were repeated over time or in experiments using a smaller number of isolates. The lesion number per leaf was significantly different depending on the isolate used for inoculation, irrespective of the plant part group. No difference was found between individual isolates when pathogenicity was assessed using a disease severity scale. No cultivar–isolate interaction was observed for lesion density when 12 isolates were tested on six cultivars, but there were some significant differences between cultivars and between isolates. Results obtained confirmed that infection by B. sorokiniana is highly variable, and very sensitive to environmental conditions. In this study no physiological specialization was observed. The fungus appeared as a continuum of isolates differing in aggressiveness. This work was conducted with isolates from a single site, and highlights the need to assess carefully the biological significance of any differences observed among B. sorokiniana isolates.     

几丁质酶基因在大肠杆菌中的表达及其酶对柱花草胶孢炭疽病原菌的？…

构建了几丁质酶基因植物表达载体,该基因大大肠杆菌中能表达３５ＫＤ的蛋白质。几丁质酶粗提高在体外对柱花草胶孢炭疽病原菌有极显著的抑菌效果。     

应用杆状病毒系统高效表达猪繁殖与呼吸综合征病毒GP5基因

将猪繁殖与呼吸综合征病毒CH_la株囊膜糖蛋白 (GP5 )基因定向亚克隆到杆状病毒转移载体pBlueBacHisB中 ,与杆状病毒线性DNA(Bac_N_BlueTMDNA)共转染昆虫细胞sf9,经过三轮蚀斑纯化后 ,获得重组杆状病毒rBac_GP5。用该重组病毒感染sf9细胞后 ,应用间接免疫荧光试验、SDS_PAGE和Westernblot检测表明 ,GP5基因在杆状病毒系统中获得了高效表达 ,表达产物因糖基化程度差异 ,表观分子量有 2 2、2 5和 30kDa三种总计约占细胞蛋白总量的 2 6 .4%。     

一株鸡大肠杆菌1型及P型菌毛蛋白结构基因的克隆

据国外发表的人病性大肠杆菌１型及Ｐ型菌毛蛋白结构基因（ｐｉｌＡ、ｐａ;ｐＡ）＿序列,设计并合成 于ＰｉｏＡ及ｐａｐＡ保守区的２对引物。经ＰＣＲ从１株带有甘露糖敏感血凝特性（ＭＳＨＡ）及甘露９糖抵抗血凝特性（ＭＲＨＡ）的鸡致病性大肠杆菌（ＨＪ２０ｅ）染色体中扩增到大小分别在６５７ｂｐ、５４ｌｂｐ的２种阳性产物。通过核酸序列测定分别确证所扩增的ＤＮＡ片段分别为ｐｉｌＡ、ｐａｐＡ基因。经酶切、连接、转化     

单抗对禽病原性大肠杆菌菌毛粘附抑制的研究

应用８株鸡病原性大肠杆菌研究了菌毛作为分型抗原和共粘附特性,以抗Ⅰ型菌毛单克隆抗体（单抗）ｂＧ５和甘露糖进行了抑制作用试验。在血凝试验中、单抗和甘露糖都能抑制血凝反应,说明这些株都属于Ⅰ型,通过以单抗或甘露糖的粘附抑制程序这些Ｅｃｏｌｉ株对鸡气管上皮都不能特异粘附。     

苜蓿种带真菌及其致病性测定

对来自我国新疆,内蒙古,吉林,陕西,甘肃,宁夏,山东,黑龙江,山西,河北和江苏等１１个省（自治区）苜蓿产区的３０个审定登记品种的３８个苜蓿种子样品进行了系统的种带真菌检测研究;测定了２１种２５个真菌分离物对苜蓿幼苗根组织的入侵性及１９种真菌对苜蓿种子萌发和幼苗生长的影响。