H1亚型猪流感病毒血凝素基因在昆虫细胞中的表达及其间接ELISA方法的初步建立

根据GenBank发表的H1亚型猪流感HA基因序列设计引物,扩增出HA基因片段.将其克隆到pFastBacGP67B杆状病毒载体上,筛选阳性重组转座载体pFastBacGP67B-H1,转化含有杆状病毒穿梭载体(bacmid)的DH10Bac感受态细胞,构建杆状病毒表达载体获得重组转座子(rBacmid-H1),在脂质体介导下转染sf9昆虫细胞,获得重组杆状病毒(rBV-H1),再感染细胞,收获目的蛋白.通过血凝试验、免疫印迹法、免疫组化分析表明该蛋白得到表达,且具有良好的生物学活性.利用表达的蛋白作为猪流感间接ELISA的抗原,初步建立H1亚型猪流感的间接ELISA检测方法,并对内蒙古、辽宁和黑龙江等地送检的93份猪血清进行了检测,阳性率为31.18%,为研制开发快速、准确、简便的H1亚型猪流感鉴别诊断试剂盒奠定基础.     

辛伐他汀对动脉粥样硬化大鼠凋亡相关基因Fas及FasL蛋白表达的影响

目的通过研究辛伐他汀对动脉粥样硬化大鼠血管壁中细胞凋亡相关基因Fas及FasL蛋白表达产物的影响,探讨其在预防动脉粥样硬化发生中的可能机制。方法复制动脉粥样硬化大鼠模型,以辛伐他汀干预,取胸主动脉,观察其斑块变化,采用免疫组化Elivision法测定动脉粥样硬化血管壁中Fas、FasL蛋白表达。结果Fas蛋白表达在实验组明显高于对照组及干预组(P<0.01,P<0.05),实验组FasL蛋白表达也明显高于对照组及干预组(P<0.05)。结论Fas及FasL基因通过促进细胞凋亡作用而诱发动脉粥样硬化过程,辛伐他汀可通过调节细胞凋亡过程发挥抗动脉粥样硬化作用。     

Fine mapping and candidate gene analysis of dense and erect panicle 3, DEP3, which confers high grain yield in rice (Oryza sativa L.)

Architecture of the rice inflorescence, which is determined mainly by the morphology, number and length of primary and secondary inflorescence branches, is an important agronomical trait. In the current study, we characterized a novel dense and erect panicle (EP) mutant, dep3, derived from the Oryza sativa ssp. japonica cultivar Hwacheong treated with N-methyl-N-nitrosourea. The panicle of the dep3 mutant remained erect from flowering to full maturation, whereas the panicle of the wild type plant began to droop after flowering. The dep3 mutation also regulated other panicle characteristics, including panicle length, grain shape and grain number per panicle. Anatomical observations revealed that the dep3 mutant had more small vascular bundles and a thicker culm than wild type plants, explaining the EP phenotype. Genetic analysis indicated that the phenotype with the dense and EP was controlled by a single recessive gene, termed dep3. The DEP3 gene was identified as the candidate via a map-based cloning approach and was predicted to encode a patatin-like phospholipase A2 (PLA2) superfamily domain-containing protein. The mutant allele gene carried a 408?bp genomic deletion within LOC_Os06g46350, which included the last 47?bp coding region of the third exon and the first 361?bp of the 3??-untranslated region. Taken together, our results indicated that the patatin-like PLA2 might play a significant role in the formation of vascular bundles, and that the dep3 mutant may provide another EP resource for rice breeding programs.     

Response surface methodology to design a selective co-enrichment broth of Escherichia coli, Salmonella spp. and Staphylococcus aureus for simultaneous detection by multiplex PCR

Escherichia coli, Salmonella spp. and Staphylococcus aureus are frequent co-visitors of contaminated foods to cause food-borne diseases. To achieve rapid detection of three organisms by multiplex PCR, a selective co-enrichment broth was considered to design using response surface methodology (RSM) in this work. NaCl, LiCl and KSCN as selective bacterial inhibitors were selected to optimize their concentrations for a matched composition of bacterial biomass with uniform amplification of three targets. Central composite design was employed to collect the data and fit the responses. Three quadratic polynomial models were derived by computer simulation. A statistical analysis was carried out to explore the effects of the variables on the composition of bacterial biomass and PCR amplification yields. In the end, a novel broth (ESS-3 broth) of NaCl 1.60%, LiCl 0.70%, KSCN 0.10% was formulated to allow co-enrichment of the target pathogens and suppress growth of some non-target pathogens. The simultaneous detection of E. coli, Salmonella spp. and S. aureus was developed on a rapid, convenient and sensitive method consisting of selective co-enrichment in ESS-3 broth, DNA extraction with the boiling method and robust test by multiplex PCR. Our work provided broader application of RSM for the simultaneous detection of other combinations of multiple pathogens.     

离子对高效液相色谱法分析黄瓜花叶病毒侵染烟草的总RNA水平变化

为研究植物病毒在RNA水平上对寄主植物的基因表达产生的影响,采用离子对高效液相色谱法,对黄瓜花叶病毒侵染烟草造成的总RNA的组成成分变化进行研究。离子对液相色谱法,是近几年才应用于对RNA进行分离、纯化和分析的一种试验技术,具有操作简单、重复性好、所需时间短等优点。选择适宜的离子对试剂,并对选定的离子对试剂正己胺/1,1,1,3,3,3-六氟-2-丙醇进行了优化,达到了较好的总RNA分离效果,并观察到健康植株和染病植株分离峰之间的差异,有差异的RNA种类还需进一步试验来验证。为研究植物病毒的致病机制提供一种新的试验方法和途径。     

云南阳宗海酵母菌种群结构及产胞外酶测试北大核心CSCD

【目的】研究阳宗海酵母菌种群结构,分析生物因子及非生物因子对酵母菌种群分布的影响;测试阳宗海酵母菌产胞外酶活性。【方法】水样用醋酸纤维素滤膜过滤,原位培养分离酵母菌;梯度稀释法分离土样和底泥样品;对分离得到的菌株进行DNA提取和测序,分析26S rDNA的D1/D2区域,并结合形态及生理生化指标进行鉴定;用产酶筛选培养基对分离得到的酵母菌进行产胞外酶活性测试。【结果】共分离得到201株酵母菌,鉴定分属于15个属48个种,其中包括10个潜在的新种;普鲁兰类酵母(Aureobasidium pullulans),库德里阿兹威氏毕赤酵母(Pichia kudriavzevii),胶红酵母(Rhodotorula mucilaginosa),Cryptococcus podzolicus是优势种;15.9%的酵母菌具有产胞外酶活性,主要是脂肪酶和淀粉酶。【结论】阳宗海酵母菌有较为丰富的多样性,人为活动对阳宗海酵母菌分布影响较大,其次浊度、电导率也是影响酵母菌种群分布的重要因素;阳宗海产胞外酶酵母菌可能参与湖泊生态系统的自然循环。     

利用数值模拟重构物种多样性格局的形成过程

生命形成的过程极其漫长, 经历了地球系统复杂的沧海桑田变化。当前人类所观察到的物种分布格局的形成除了由物种本身特征决定外, 还受到环境变化、人类活动以及各种随机事件的影响。受限于实验条件、时间、经费、人力等诸多因素, 我们尚无法完整地观察并记录到物种多样性形成的全过程, 只能通过片段化数据来推测该过程。信息科学中包括数值模拟在内的仿真技术以其高效、可控及全过程记录等优势, 能从某种程度上解决物种多样性格局形成过程中的部分数据黑箱问题。本文介绍了数值模拟的概念和工作原理及在物种多样性研究中应用的特点, 列举了物种生态位、扩散模式、种间互作及物种分布应对气候变化等方面的数值模拟研究, 基于已有研究系统地介绍了如何综合上述数值模拟研究构建虚拟物种、气候和场景来解释物种多样性的形成与维持机制, 并阐述了数值模拟在物种多样性研究中的优缺点及应用前景。     

神经递质的可视化荧光检测技术研究进展

神经递质是神经系统中至关重要的组成部分,神经递质释放的时间和空间变化是神经网络中信息处理的核心,可视化监测神经递质的生物传感器是探究各类生理和病理活动的重要工具。文中综述了近年来具有较高时间和空间分辨率的监测神经递质时空分布变化技术的研究进展,介绍了对谷氨酸、多巴胺、γ-氨基丁酸和乙酰胆碱这4类重要的神经递质的检测方法,并归纳总结了各类检测方法的基本原理和优缺点,为设计具有高时空分辨率的神经递质传感器提供一个较为系统的参考。     

紫胶林-农田复合生态系统地表甲虫多样性——以云南绿春为例

为揭示紫胶林-农田复合生态系统地表甲虫多样性, 于2006-2007年在云南省绿春县牛孔乡采用陷阱法调查了天然紫胶林、人工紫胶林和旱地的地表甲虫群落。共采集标本1 678头, 分别隶属于24科120种, 其中步甲科(Carabidae)和金龟科(Scarabaeidae)种类最丰富, 均占全部种类的12.50%。拟步甲科(Tenebrionidae)个体数量最丰富, 占个体总数的64.48%; 金龟科次之, 占个体总数的17.58%。大多数科的物种数和个体数在不同土地利用生境中的分布没有显著差异, 而步甲科、隐翅甲科(Staphylinidae)、叩甲科(Elateridae)、拟步甲科、瓢虫科(Coccinellidae)和小蠧科(Scolytidae)在不同土地利用生境中的分布有显著差异。天然紫胶林地表甲虫个体数量最少, 物种较丰富, 优势度最低, 多样性最高; 人工紫胶林个体数和物种数均丰富, 优势度和多样性居中; 旱地个体数量最多, 物种最贫乏, 优势度最高, 多样性最低。种级水平的聚类分析体现出人工紫胶林与旱地之间在种类组成上距离较近; 而科级水平的聚类反映出人工紫胶林和天然紫胶林更接近。结果提示, 紫胶林-农田复合生态系统具有区域内土地利用方式多样化的特点; 天然紫胶林在维持地表甲虫多样性水平上具有重要作用, 而人工紫胶林虽具有积极作用, 但仍需进一步恢复。