应用寡核苷酸微阵列检测基因突变     

祝骥  段俊  梁承邺 《广西农业生物科学》2003,22(3):221-224

寡核苷酸阵列技术是一种高度集成化、快速的基因分析手段，已广泛用于基因突变检测、序列测定、基因表达分析、疾病诊断等。本文主要介绍应用该技术进行基因突变检测的三种方法：杂交信号增益法、杂交信号消减法与微测序分析。三种方法各有优势，通过选择适当的方法，将为基因突变检测提供更佳的分析途径。  相似文献   

五种鸡呼吸道传染病基因芯片诊断技术的研究     

陈凤梅  吴时友  尹燕博  牛钟相 《家畜生态》2006,(2)

用分子克隆方法获得新城疫病毒、禽流感病毒、传染性支气管炎病毒、传染性喉气管炎病毒,鸡毒支原体各一段高度保守基因片段作为探针,用芯片点样仪点样到硝酸纤维膜上,制成检测基因芯片;提取样品中的RNA进行反转录,然后在PCR过程中用生物素标记,将PCR产物滴加到芯片上进行特异性杂交,对杂交结果进行扫描分析,可同时对上述5种常见禽呼吸道疾病作出诊断。此方法与PCR检测方法结果符合率在95%以上,其检出率比临床分离法要高30%以上。  相似文献   

Gene expression profiles in bovine granulocytes reflect the aberration of liver functions     

Keiichiro Kizaki  Tomomi Kageyama  Noriyuki Toji  Katsuo Koshi  Kouya Sasaki  Norio Yamagishi  Toshina Ishiguro‐Oonuma  Toru Takahashi  Kazuyoshi Hashizume 《Animal Science Journal》2020,91(1)

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禽流感病毒NP基因的真核表达及抗体检测芯片的建立     

赵玉辉  王馥梅  包红梅  孙晓东  路冰  熊永忠  陈化兰  王秀荣 《中国预防兽医学报》2011,33(5)

为建立一种检测A型禽流感病毒(AIV)抗体的蛋白芯片,本研究以AIV总RNA为模板,RT-PCR方法扩增NP基因,与真核表达载体pFastBacHTa连接,并在昆虫细胞中表达.重组蛋白经纯化及western blot鉴定,点样于醛基包被的载玻片上,制备检测AIV抗体的蛋白芯片.确定芯片反应最佳条件为:点样浓度2 mg/mL,固定24 h,1%BSA进行封闭,一抗稀释度1:100,二抗稀释度1:2000.利用蛋白芯片分别对15个亚型流感病毒免疫血清、SPF鸡血清、抗新城疫病毒血清、抗法氏囊病毒血清和现地血清样品进行检测,通过与血凝抑制试验比较,表明建立的蛋白芯片方法具有良好的灵敏性和特异性,为AIV抗体检测及制备检测AIV不同亚型或多种病原抗体的蛋白芯片提供方法.  相似文献   

Development of a low-density DNA microarray for diagnosis of target-site mutations of pyrethroid and organophosphate resistance mutations in the whitefly Bemisia tabaci     

Chung IH  Kang S  Kim YR  Kim JH  Jung JW  Lee S  Lee SH  Hwang SY 《Pest management science》2011,67(12):1541-1548

BACKGROUND: Rapid and accurate detection of mutations related to insecticide resistance is essential for development of resistance management strategies to support sustainable agriculture. The M918V, L925I and T929V mutations of the voltage‐gated sodium channel gene (vgsc) and the F392W mutation of the acetylcholinesterase I gene (ace1) are reportedly associated with resistance to pyrethroids and organophosphates, respectively, in Bemisia tabaci. In order to detect known base substitutions in the ace1 and vgsc genes, a low‐density microarray with an allele‐specific probe was developed. RESULTS: Specific regions of the ace1 and vgsc gene mutations were amplified by multiplex asymmetrical PCR using Cy3‐labelled primers, and then the PCR products were hybridised on the microarray. After analysing the probe signal data, the microarray containing 12 allele‐specific probes produced a unique pattern of probe signals for field DNA samples of B. tabaci. To determine the optimal cut‐off value of each probe, receiver operating characteristic (ROC) curve analysis was conducted using SPSS. Among 60 individual samples, microarray data for 57 samples were consistent with direct sequencing data. CONCLUSION: Although many molecular detection methods have been employed to monitor insecticide resistance, the present microarray provides rapid and accurate identification of target mutations in B. tabaci for resistance management. Copyright © 2011 Society of Chemical Industry  相似文献   

用聚赖氨酸包被国产载玻片制作DNA芯片     

徐景升  贾锡伟  邹志华  王艺磊  张木清  陈如凯 《福建农林大学学报(自然科学版)》2004,33(4):485-488

用多聚赖氨酸对国产载玻片进行包被处理,在不同的固定条件下,比较其与进口多聚赖氨酸载玻片对大片段DNA和3′端氨基修饰寡核苷酸的固定效果.结果表明,在DNA固定率上,自行包被处理的国产载玻片与进口多聚赖氨酸载玻片没有显著差异;紫外交联对大片段DNA固定率影响显著,但对氨基修饰的寡核苷酸影响不显著;3种点样液中,进口点样液MSS点样效果最好,其次为3×SSC,第三是水.氨基修饰的寡核苷酸可以很好地固定在聚赖氨酸包被的载玻片上.  相似文献   

基因芯片技术鉴定棉铃虫胁迫后棉花差异表达基因   总被引：1，自引：0，他引：1  

武娟  齐放军  田文华  韩榕  张永军  郭予元 《农业生物技术学报》2011,19(5)

棉花在受到植食性昆虫为害后基因表达会发生变化,筛选鉴定这些调控基因有助于解析昆虫诱导棉花抗虫性的分子机制。本研究以棉花(Gossypium spp.)(中12)为实验材料,分别从正常生长条件下的棉花(对照)和棉铃虫(Helicoverpa armigera Hübner)取食胁迫6、12、24和48h的棉花叶片(处理)中提取总RNA,采用Affymetrix棉花基因芯片进行了基因表达谱分析。结果表明,棉铃虫取食6h时,差异表达基因共有4109个(占28%),其中上调1917个,下调2192个;棉铃虫取食12h时,差异表达基因共有2605个(占18%),其中上调1326个,下调1279个;棉铃虫取食24h时,差异表达基因共有3213个(占22%),其中上调1424个,下调1789个;棉铃虫取食48h时,差异表达基因共有2763个(占19%),其中上调1450个,下调1313个。进行生物信息学分析后发现,这些基因功能涉及氧化应激响应、防御响应、信号转导、转录调控、黄酮类生物合成、萜类化合物合成与代谢以及源于多种氨基酸的罗勒生物碱生物合成等多个方面。另外,从芯片结果中鉴定获得调控棉花特异性挥发物的相关基因,发现法呢...  相似文献   

蛋白芯片在蛋白质组学中的应用     

于晓波  郝艳红  许丹科  李庆章 《东北农业大学学报》2006,37(2):276-282

最近几年,蛋白芯片技术发展很快并在蛋白质组学通量化和系统化的研究中扮演越来越重要的角色,展现出良好发展前景。为此,文章对蛋白芯片的主要原理和类型以及近几年蛋白芯片在蛋白质组学中的应用,尤其是在蛋白表达谱图和蛋白质相互作用谱图中的应用进行了综述。  相似文献   

Characterization of NtSTOP1-regulating genes in tobacco under aluminum stress     

Hiroki Ito  Yuriko Kobayashi  Yoshiharu Y. Yamamoto 《Soil Science and Plant Nutrition》2013,59(3):251-258

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cDNA microarray analysis of interleukin-1β-induced Japanese flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis> kidney cells     

Dhanwanthari?Emmadi  Akiko?Iwahori  Ikuo?Hirono  Takashi?AokiEmail author 《Fisheries Science》2005,71(3):519-530

ABSTRACT:   Interleukin-1β (IL-1β) cDNA of Japanese flounder was found to consist of 1329 bp, encoded 247 amino acid residues. Among the fish IL-1β in the databases, the one with the highest identity of Japanese flounder IL-1β was that of seabass (62% identity). The expression of IL-1β was induced by treatment with concanavalin A (ConA)/phorbol myristate acetate (PMA) and lipopolysaccharide. The copy number of IL-1β mRNA was increased 30-fold after stimulation with ConA/PMA. Of 871 cDNA on a microarray, 93 genes (10.7%) were up-regulated or down-regulated by IL-1β at 1, 3 and 7 days post-injection. The induced gene expression was highest on day 1 followed by day 3 and day 7. A total of 7% of known and 3.7% of unknown genes of the 871 tested genes were differentially expressed. Of the genes tested, 7.4% were up-regulated and 3.3% were down-regulated. Cytokine genes such as tumor necrosis factor, granulocyte colony stimulating factor and chemokine receptor A were induced in response to IL-1β. Cell surface antigens such as IgM, MHC class I and CD20 receptor were up-regulated. Signal transduction genes such as Toll-like receptor 1 and SH3P2 were also up-regulated. The glucocorticoid receptor and cAMP early repressor were down-regulated in our microarray analysis.  相似文献