快速高效堆肥处理城市污泥微生物多样性研究

采用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术对城市污泥堆肥工艺中微生物种群多样性进行初步研究，结果表明污泥中的微生物种群有显著的改变，同时对堆肥后污泥中的微生物进行了尝试性探讨，为城市污泥微生态和高效堆肥工艺的研究提供了新的实验依据。     

恩施悬棺人骨mtDNA初步分析

本文以恩施宋代悬棺的骨骼或牙齿样品为对象,经过一系列的处理,获得并扩增了mtDNA的HVSI区的部分序列,通过几个实验初步认定是古DNA.同时通过与部分现代人mtDNA数据的比较参照,对所研究样品单倍群归属进行了初步确定,发现他们主要属于南方类型,但也可能有北方成分.     

Detection of Grass Carp Hemorrhage Virus （GCHV） from Vietnam and Comparison with GCHV Strain from China

Grass carp plays an important role in small-scale aquaculture in Vietnam. However, a severe disease, known in Vietnam as “Red Spot Disease“, is causing significant economic loss in grass carp aquaculture. In this study, the tissue samples isolated from the grass carp with Red Spot Disease in Vietnam are investigated and eomparied with the control GCHV isolated in China by experimental infection, culture cell infection, serological cross reactivity, and RT-PCR amplifica-tion. Infected grass carp exhibits hemorrhage symptoms about 5 days after experimental injection with GCHV-V (Vietnam) strain. The symptoms and lethality induced by the GCHV-V strain are identical to that induced by the Chinese GCHV-9014 strain. The Chinese GCHV-873 strain in-duces typical cytopathogenic effects in 4 cell lines, such as CIK, CAB, FHM and GCO, from the6 fish cell lines examined. No cytopathogenic effects are observed in all the 6 examined cell lines,including CAB, FHM, CIK, EPC, CCO and G(X), infected by the GCHV-V strain and GCHV-9014 strain. Immunodiffusion assays demonstrate an obvious cross-reactivity among three GCHVstrains. Precipitin lines are clearly observed not only between the anti-GCHV-873 serum and thetwo strains GCHV-873 and GCHV-9014, but also between the anti-GCHV-873 serum and the GCHV-V strain. GCHV can be detected by immunodiffusion assays after three generations of blind propagations in the cell lines inoculated by GCHV-V strain. This implicates that GCHV-Vviruses have been replicated and amplified despite there being no cytopathogenic eifects observed in these examined cell lines. Three genome segments of GCHV, including S8, S9 and S10, are amplified by three sets of PCR primers designed according to the segment sequences published re-cently. The Q8fp and Q8rp primer set specific for genome segment S8 amplifies a 955 bp frag-ment from the extracted sample of diseased fish with Red Spot Disease, and the fragment size is i-dentical to that amplified by the same primer set from control GCHV-873 strain. Simultaneously,the Q9fp and Q9rp primer set specific for genome segment S9 generates a same 635 bp product,and the Q10fp and Q10rp primer set specific for genome segment S10 produces a same 697 bp fragment from both template samples of diseased fish with Red Spot Disease and control GCHV-873 strain. The RT-PCR amplification and corresponding size comparison data indicate that the three GCHV-V genome segments extracted from the diseased grass carp with Red Spot Disease in Vietnam should be identical to that in control GCHV-873 strain from China. The data confirm that the causative agent of grass carp Red Spot Disease in Vietnam is a virus, and the virus is closely similar to GCHV strain in China.     

克隆胃粘膜细胞癌变相关基因的思路

采用ｍＲＮＡ和ＡＰ－ＰＣＲ等技术克隆胃粘膜细胞癌变相关基因在技术上是可行的。胃粘膜细胞癌变相关基因的获得，在阐明胃癌发生机制及胃癌的早期诊断和临床治疗方面，具有重要意义和广阔的应用前景。     

聚合酶链式反应仪基座的热阻建模与分析

对聚合酶链式反应仪(PCR)基座中温度变化规律进行数学建模,求出其过程热阻值,并利用该热阻值对类似模型的温度场进行仿真计算,从而快速地得出考虑若干关键参数的复杂模型的温度场.计算结果显示,PCR仪基座的过程热阻与其自身的导热热阻存在某种关系,进而提出一个对于该关系的假设和数学描述,最后对该假设应用的可行性进行数值计算证明.     

Bst DNA聚合酶的纯化及等温扩增检测体系的构建

Bst DNA聚合酶是等温扩增反应中的一个关键酶,但由于专利限制,目前国内实验室所使用的Bst DNA聚合酶大多需要从国外进口,价格昂贵且长途运输影响酶活性,因此亟需自主生产出Bst DNA聚合酶应用于等温扩增检测技术.通过构建Bst Fragement的原核表达质粒,采用原核蛋白表达系统和His-Tag亲和纯化手段,制备成本低,活性高,特异性强的Bst DNA聚合酶.同时,针对特定的模板,对自制Bst DNA聚合酶的扩增反应进行了进一步优化,用不同的产物鉴定方法对扩增效果进行检测,并最终筛选出该酶最优的扩增反应条件为60 mM K+、30 mM(NH4)2SO4、pH为9.0.最后,验证了Bst DNA聚合酶能用于快速准确检测肺炎支原体、肺炎衣原体,降低了相关病原体核酸检测的成本,为之后核酸检测在基层医疗的更广泛应用提供了条件.     

Multiplex chimeric primers used in a tetra-primer amplification refractory mutation system PCR to detect two single nucleotide polymorphisms associated with ovarian cancer

目的:建立一种简单快速的方法,可同时检测两个卵巢癌相关单核苷酸多态性位点rs608995和rs749292.方法:采用多重嵌合引物策略改进四引物扩增受阻突变体系聚合酶链式反应(PCR),通过设计rs608995和rs749292的特异性嵌合引物,经单管一次PCR,琼脂糖电泳分析产物长度.结果:对46份全血样本,电泳结果均与测序一致,可准确分型.结论:经多重嵌合引物策略改进的四引物扩增受阻突变体系PCR可简便、快速、准确的获得2个位点的分型结果.     

基因也存在“生理周期”

人体活动有着24小时的节律周期。以前认为,只有少数基因和蛋白质是周期性地打开、关闭,但美国科学家发现,身体各个器官的数千个基因,每天的起伏变化也都是可预测的,它们的活动周期则受多种复杂方式的控制。了解基因在一天中如何周期性地开关,是掌握许多生理功能的关键,包括睡眠和新陈代谢。霍华德?休斯医学院研究人员、得克萨斯大学西北医学中心的约瑟夫·塔卡哈斯在20世纪90年代发现了节律基因及其蛋白质产物,他和其他研究人员确定了该基因为CLOCK,并发现其他两种蛋白BMAL1和NPAS2,能在白天与基因结合激活它们,另外4个节     

多重聚合酶链反应检测污水中产气荚膜梭菌

根据产气荚膜梭菌产生4种主要毒素（α、β、ε、τ-毒素）可分为A～E5种菌型。依据产气荚膜梭菌的5种毒素基因的序列，设计5对PCR引物，建立多重聚合酶链式反应（PCR）方法，以快速区分5种类型的产气荚膜梭菌，68份环境样品（生活污水、屠宰场污水）的常规检测和基因分型共检出55株产气荚膜梭菌，其中A型产气荚膜梭菌51株，占总数的90%以上，B，C，D3种类型只占5%。试验未检出E型产气荚膜梭菌，所有的分离株未发现带有肠毒素，结果表明：目前污水中主要存在的菌型为A型菌，其他菌型的产气荚膜梭菌很少，而多数是非致病性产气荚膜梭菌。     

细胞与病毒中的RNA依赖的RNA聚合酶

就细胞与病毒的RdRP的结构及其功能，以丙型肝炎病毒的NS5B蛋白和RNA干扰系统为例，介绍了RdRP的研究进展．