低钾胁迫对烟草幼苗碳氮代谢基因表达谱的影响

为探索烟草响应低钾胁迫的分子机制,对K326烟草幼苗进行了0、6、12和24 h的低钾胁迫处理,并利用定制的烟草全基因组芯片,分析了烟草幼苗基因表达谱的变化。结果表明,与0 h相比,6、12和24 h的3个时间点差异表达在2倍以上(p<0.05)的基因总数为3790个。从功能方面看,这些差异表达基因广泛参与了烟草的各种生物学过程,包括抗氧化活性、应激反应、运输活性、发育过程、催化活性、生物调节、代谢过程等。其中包括了硝酸还原酶基因NIA2等10个参与烟草氮代谢的基因,以及UGP等44个参与烟草碳水化合物与糖代谢的基因。本研究结果说明,低钾胁迫对烟草的基因表达产生了广泛的影响,进而可能影响到烟草的碳氮等基础代谢。     

基因表达谱技术分析棒曲霉毒素肾细胞毒性的机制

本文研究了食品污染物棒曲霉毒素诱导人胚肾细胞损伤过程中基因表达谱动态变化。5μM PAT作用HEK293细胞3、24h,应用数字基因表达谱(DGE)技术筛选差异表达基因,分析基因富集的GO功能,KEGG信号通路及Pathway信号途径分析,并通过定量PCR对部分差异表达基因进行验证。分析发现,在3 h和24 h受PAT诱导的差异表达基因分别为79个和261个,其中上调表达基因分别为71和192个,下调表达基因分别为8和69个。PAT主要诱导了细胞凋亡基因的差异表达,RNA降解差异基因全部表达上调,RNA合成差异基因表达下调,泛素介导的蛋白降解大部分差异基因上调,MAPK信号通路主要参与PAT毒性的下游进程,其相关差异基因大部分上调。DGE技术为我们在分子水平上提供了PAT胚肾细胞损伤机制的众多线索,为相关基因生物学效应研究奠定基础。     

大蒜喂养对小鼠脾脏基因表达的影响

大蒜是一种常用的调味剂,它在抗动脉粥样硬化、抗增殖和降血脂方面的效应已为人所知,然而其作为膳食补充剂如何影响机体的作用机制仍未清楚。因此,分析了以大蒜喂养与对照(添加纤维素)的小鼠基因芯片表达谱,从全基因组水平探讨大蒜对小鼠生理的影响。结果发现:表达下调的基因多为胰腺消化酶及蛋白酶,上调的基因则和血红素合成、红细胞生成等生物功能相关。这些基因的表达变化可能是大蒜具有多种生理功效的原因。     

秦巴不同生态区烤烟基因表达谱分析

为了揭示不同生态区烤烟风格特色形成的机理,采用Agilent 烟草寡聚芯片技术,对商洛市洛南县和宝鸡市陇县(对照)两个生态区烤烟打顶后15 d 同一品种(秦烟96)中部叶的基因表达谱进行了初步分析。结果表明:与陇县相比,在44 000 个基因中洛南烟区烟叶有2 058 个基因差异表达,其中上调基因962 个,占46.74%;下调基因1 096 个,占53.26%。按功能富集分类,这些差异基因主要富集在代谢、响应外界刺激及生理调控等生物学过程,大部分位于胞外区与细胞内膜系统。其中洛南烟区碳水化合物合成代谢较为活跃,陇县烟区萜类化合物代谢、苯丙烷代谢、芳香族化合物合成代谢较为活跃。基因代谢通路分析表明,细胞信号转导、转录因子及参与植物非生物胁迫相关基因在两个烟草种植区均为高表达。      

高通量测序在动物检疫中的应用进展

面对无法预测、爆发突然及传播迅速的疫情,快速获知病原信息并真实地反映样品及环境中病原的多样性资料是各国卫生部门及口岸预警的核心工作之一。应用高通量测序技术全面监控研究复杂病原样品,是协助寻找传染溯源、防控动物疫病可行性的一条重要思路。目前,高通量测序在动物检疫上的应用与传统的病原研究方式最大的区别在于把病原微生物看成一个整体,摆脱了对单一病原分离培养的步骤,直接对样品基质中所有病原进行研究。随着测序技术和数据处理分析能力的飞速发展,以高通量测序为基础的宏基因组学研究已渗透到各个领域,在畜产品、养殖环境等样本检测方面显示了重要价值。本文对高通量测序的研究方法及在检验检疫上的应用趋势进行了综述。     

荧光探针在高通量筛选技术上的应用进展

荧光探针是一种分析检测工具,针对特定的化学物质可以将微观的识别信息转化为宏观的荧光信号,具有高灵敏度、高选择性、检测体系干扰小、操作简便等优点.基于荧光探针的优势,其在高通量筛选技术领域有着重要的应用.该文主要综述了荧光探针的起源、作用机理以及国内外关于荧光探针在高通量筛选技术上的应用进展,旨在为拓宽荧光探针技术的应用...     

烟草甲气味受体基因的鉴定和组织表达谱

为深化对烟草甲嗅觉感受系统的认识,促进其行为调控剂的开发,利用BLASTX在线工具和系统进化分析等生物信息学方法从烟草甲触角转录组数据中挖掘鉴定烟草甲气味受体基因。研究结果表明:①从烟草甲触角转录组中共鉴定出9个气味受体基因（LerOR1~LerOR8、LserOrco）。②系统进化树中,烟草甲的LserOR6与光肩星天牛AglaOR22聚在同一个分支,表明它们之间的进化关系更接近。③烟草甲的2个气味受体基因（LserOR5和LserOrco）在触角中表达量相对较高。      

外源质粒DNA对小鼠肝脏基因表达谱的影响

研究了外源质粒DNA经胃肠道吸收可能对肝脏产生的作用机制。给Balb／c小鼠灌胃质粒pcDNA3 200μg，在灌胃后4h分离肝脏，提取肝脏的总RNA。利用寡核苷酸芯片对灌胃质粒pcDNA3后的Balb／c小鼠肝脏进行基因表达谱研究。结果发现17664个基因中，表达上调100条，表达下调41条。按基因功能分类，表达上调的基因可分为免疫应答基因、转录因子基因、信号转导基因、转运相关基因及代谢相关基因等；表达下调的基因主要为脂质代谢基因。灌胃外源质粒DNA后，肝脏组织主要表现为急性时相反应的加强、免疫反应的活化、细胞信号通路的活化以及脂质代谢途径的抑制。表明外源质粒DNA通过胃肠道途径可广泛调控肝脏的基因表达。     

高通量测序技术在乳制品研究中的应用

高通量测序技术以一次并行对几十万到几百万条DNA分子进行序列测定为标志,目前已广泛应用于各个领域,对整个生命科学的发展起到极大的推动作用。本文结合前期开展的一些工作,综述高通量测序技术在乳制品微生物基因组和生物多样性中的应用情况。     

高通量花生转录组分析系统的构建与应用

采用PC机为硬件平台,基于Linux操作系统,以Perl语言作为主要的程序编辑语言,整合SeqClean、TGICL、RepeatMasker、Blast、Bioperl等软件或模块,构建自动化、高通量的转录组分析系统。整个分析系统以三个自编的Perl脚本为主程序,其中以assemble．pl为主程序的序列组装,以annotFun．pl为主程序的功能注释和以annot—GO．pl为主程序的功能分类。通过分析粤油20在叶斑病诱导下获得的8328个叶片EST,进一步验证该系统的稳定性和可靠性。本文构建的花生转录组分析系统通过三个主程序能够自动、简洁、高通量地完成花生转录组数据的组装、注释与分类,为花生功能基因组学研究提供有价值的生物信息,也为其他生物信息平台的构建提供借鉴。     

Global gene expression profiling of androgen disruption in Qurt strain medaka

Androgen disrupting chemicals (ADCs) are endocrine disrupting chemicals (EDCs) that mimic or antagonize the effect of physiological androgens. Microarray-based detection of altered gene expression can be used as a biomarker of EDC exposure. Therefore, the purpose of this study was to identify and compare gene expression profiles of the androgen 11-ketotestosterone (11-KT), the antiandrogen flutamide (FLU), and the antiandrogenic fungicide vinclozolin (VIN), on Qurt medaka (Oryzias latipes). Biologically effective concentrations for 11-KT (100 microg/L), VIN (100 microg/L), and FLU (1000 microg/L) determined in range-finding studies were used for exposures. The oligonucleotide microarray included 9379 probes for EDC-affected genes, medaka cDNAs, sequences from the medaka genome project, and the UniGene database. We found that treatment with FLU, VIN, and 11-KT caused significant (false discovery rate = 0.01) differential expression of at least 87, 82, and 578 genes, respectively. Two sets of responsive genes are associated to vertebrate sex differentiation and growth, and 50 genes were useful in discriminating between ADC classes. The discriminating capacity was confirmed by a remarkable similarity of the antiandrogenic expression profiles of VIN and FLU, which were distinct from the androgenic profile of 11-KT. Gene expression profiles characterized in this study allow for reliable screening of ADC activity.     

痕量样本高通量测序文库构建

目的 通过调整建库过程中的测序接头加入量、片段选择的执行与否、文库扩增循环数等条件,建立一种基于Illumina测序平台的高效稳定的痕量样本高通量文库构建方法.方法 采用不同类型的样本,设计不同起始DNA量(最低至0.1 ng)的文库构建策略,主要对接头使用量、扩增循环数等进行优化,并通过数据分析评估文库质量.结果 对...     

Biological responses of ligament fibroblasts and gene expression profiling on micropatterned silicone substrates subjected to mechanical stimuli

In this study, ligament fibroblasts were cultivated on micropatterned silicone substrates and subjected to cyclic stretching to simulate the in vivo biomechanical environment during ligament healing. Without stretching, ligament fibroblasts were aligned parallel to the microgrooves on the silicone substrate surface. However, we previously reported that uniaxial cyclic stretching induces alignment perpendicular to the stretching axis. With stretching on a microgrooved surface, cell proliferation and collagen production were greatly enhanced. The exact functions of the micropatterned surface and mechanical stimuli are unknown. Therefore, in gene expression microarray experiments, genes whose expression is inhibited by subculture from passage 0 (P0) to passage 8 (P8) and enhanced by micropatterning and stretching were sought out. The following six genes were selected: MGP, GADD45A, UNC5B, TGFB1, COL4A1, and COL4A2. The selected genes play fundamental roles in cell proliferation, differentiation, apoptosis, and structural maintenance. On the basis of the obtained gene expression profiles, we identified candidate genes that might be involved in responses to a micropatterned surface and mechanical stretching.     

食品中违禁色素种类及高通量检测技术进展

工业染料是食品中常见的违法添加色素,是食品中常见的非法添加物。本文根据工业染料的化学性质,将食品中易违法添加的近40种违禁色素分为酸性,中性和碱性,对它们的理化性质、易非法添加的食品种类、毒性危害与法规限制,以及多种禁用色素多重高通量分析测试方法的前处理、线性范围、检出限和回收率等方面做出综述,并分析了各种检测技术的优缺点和适用性。作者认为非食用色素检测技术将趋于利用液相色谱串联质谱的高通量筛选和利用免疫学或电化学的快速检测方向发展,检测方法将更快速、环保、低成本、高自动化程度,并会针对不同的检测需求制定相应的检测方法,分别满足实验室确认分析和现场快速筛查的需求。     

Global gene expression profiling in larval zebrafish exposed to microcystin-LR and microcystis reveals endocrine disrupting effects of Cyanobacteria

Microcystis blooms occur worldwide and threaten aquatic ecosystems and human health. Sublethal effects on early developmental stages of fish are largely unknown, and research has mainly focused on microcystin toxins (such as MC-LR) rather than Microcystis cells. We exposed (96 h) zebrafish larvae to purified MC-LR (0-1000 μg/L) or lyophilized Microcystis aeruginosa containing 4.5 μg/L MC-LR and evaluated changes in global gene expression (Affymetrix GeneChip zebrafish genome arrays). Significant changes in gene expression (≥ 1.7-fold change, p < 0.0001) were determined with Rosetta Resolver 7.0, and ontology analysis was conducted with the DAVID bioinformatics tool. The number of differentially expressed genes relative to control increased with MC-LR concentration and included genes related to known mechanisms of action for MC-LR in mammals and older life stages of fish, as well as genes unique to larval zebrafish. Up-regulation of vitellogenin genes (vtg) (19.2-fold to >100-fold on arrays; 619.3-fold confirmed by quantitative PCR) was observed in Microcystis-exposed larvae but not in larvae exposed to MC-LR. Up-regulation of vtg indicates exposure to estrogenic substance(s) and suggests that Microcystis may be a natural source of environmental estrogens. Concerns about effects of Microcystis blooms may extend beyond those associated with the microcystin toxin.     

Cloning and high expression of catalase gene from bacillus sp. TE124

An efficient expression system for producing catalase in Bacillus was developed. A catalase was purified from Bacillus sp. TE124 and the catalase gene was cloned by plaque hybridization with a probe constructed from the N-terminal amino acid sequence of the enzyme. The gene, containing an open reading frame of 1452 bp, was subcloned into pHY300PLK for self-cloning into the organism. As a result, the production of catalase increased 20-fold over that of the parent strain.     

Selection of causal gene sets for lymphoma prognostication from expression profiling and construction of prognostic fuzzy neural network models

To assess the response of lymphomas to chemotherapy, gene expression profiling data from DNA microarrays were analyzed using the fuzzy neural network (FNN) modeling method. We used the FNN modeling method to produce 10 noninferior models. Using these models, we were able to predict diffuse large B-cell lymphoma (DLBCL) patient outcome with 93% accuracy. Of the 37 genes in the 10 models, 13 genes were repeatedly selected, indicating that these genes are important for prognostication. On Kaplan-Meier plots of overall survival, patients predicted by the FNN model to be cured survived significantly longer than those predicted to be refractory (P<0.0001), indicating that the FNN could successfully identify patients with a relatively poor prognosis among low-clinical-risk patients. The FNN modeling method presented here is able to precisely extract significant biological markers affecting prognosis.