We used cDNA microarrays containing ～9,000 unigenes to identify 486 salt responsive expressed sequence tags (ESTs) (representing ～450 unigenes) in shoots of the highly salt-tolerant rice variety, Nona Bokra (Oryza s...We used cDNA microarrays containing ～9,000 unigenes to identify 486 salt responsive expressed sequence tags (ESTs) (representing ～450 unigenes) in shoots of the highly salt-tolerant rice variety, Nona Bokra (Oryza sativa L. Ssp.Indica pv. Nona). Some of the genes identified in this study had previously been associated with salt stress. Howeverthe majority were novel, indicating that there is a great number of genes that are induced by salt exposure. Analysis of the salt stress expression profile data of Nona provided clues regarding some putative cellular and molecular processes that are undertaken by this tolerant rice variety in response to salt stress. Namely, we found that multiple transcription factors were induced during the initial salt response of shoots. Many genes whose encoded proteins are implicated in detoxification, protectant and transport were rapidly induced. Genes supporting photosynthesis were repressed and those supporting carbohydrate metabolism were altered. Commonality among the genes induced by salt exposure with those induced during senescence and biotic stress responses suggests that there are shared signaling pathways among these processes. We further compared the transcriptome changes of the salt-sensitive cultivar, IR28, with that of Nona rice. Many genes that are salt responsive in Nona were found to be differentially regulated in IR28. This study identified a large number of candidate functional genes that appear to be involved in salt tolerance and further examination of these genes may enable the molecular basis of salt tolerance to be elucidated.展开更多
【目的】探讨菜心抗虫性的分子机制,为抗虫品种选育和抗虫基因挖掘提供理论支持和中间材料。【方法】选用抗小菜蛾品种65和感小菜蛾品种69菜心为亲本构建F2作图群体,利用cDNA-AFLP标记构建转录图谱、分析差异基因表达,筛选和克隆抗虫相...【目的】探讨菜心抗虫性的分子机制,为抗虫品种选育和抗虫基因挖掘提供理论支持和中间材料。【方法】选用抗小菜蛾品种65和感小菜蛾品种69菜心为亲本构建F2作图群体,利用cDNA-AFLP标记构建转录图谱、分析差异基因表达,筛选和克隆抗虫相关转录衍生片段。【结果】14对引物组合共扩增出330条扩增带,其中238条为多态性带,多态性条带比例为56.00%~100.00%,平均为73.86%,平均每对引物扩增24条带,多态性条带17个。构建的cDNA-AFLP转录图谱由5个连锁群组成,193个标记未能定位在连锁群上,42个位点定位在5个连锁群上,总长度为575.869 c M,平均图距为115.1738 c M。每个连锁群上的标记数为2~23个,平均为8个。5个连锁群的LOD值变化范围为9.923~12.71,平均为13.207。A1连锁群上的分子标记分布不均匀,出现成簇分布现象和出现了5个较大的断裂。共有18个偏分离位点,偏分离比例为42.86%。在330个位点中,异常分离位点103个,占总位点数的34.24%。对克隆测序成功的10个转录衍生片段分析表明,1个TDF5-1-1与防御相关基因GW775571.1具有高度的序列同源性,8个TDF与大白菜花蕾败育相关基因GR308173.1高度同源,1个与功能未知基因EX106546.1高度的序列同源。【结论】cDNA-AFLP标记能可初步从转录组水平上揭示抗虫菜心群体的连锁遗传规律及抗虫分子机制,其抗性相关基因可进一步用于小菜蛾抗性育种。展开更多
番茄果实富含的次生物质严重干扰其总RNA的提取,本试验采用4种方法对番茄果实总RNA进行提取,筛选出一种适合于番茄后续的cDNA-扩增片段长度多态性(amplified fragment length polymorphism,AFLP)分析的高质量RNA提取方法,该方法排除了...番茄果实富含的次生物质严重干扰其总RNA的提取,本试验采用4种方法对番茄果实总RNA进行提取,筛选出一种适合于番茄后续的cDNA-扩增片段长度多态性(amplified fragment length polymorphism,AFLP)分析的高质量RNA提取方法,该方法排除了蛋白、多糖、以及多酚等次生物质的干扰,总RNA得率高、完整性好、纯度高。通过cDNA-AFLP分析方法的检测,得到了一组条带清晰和多态性丰富的图谱。展开更多
目前肿瘤癌原125（CA125）检测被广泛应用于临床卵巢癌的诊断，但由于子宫内膜异位症、盆腔炎症等也会导致CA125升高，因此，需要一种敏感性和特异性较强的标志物来提高卵巢癌的诊断水平。人附睾蛋白4（human epididymis protein 4，HF...目前肿瘤癌原125（CA125）检测被广泛应用于临床卵巢癌的诊断，但由于子宫内膜异位症、盆腔炎症等也会导致CA125升高，因此，需要一种敏感性和特异性较强的标志物来提高卵巢癌的诊断水平。人附睾蛋白4（human epididymis protein 4，HFA）基因最早是从人附睾上皮细胞中发现，1991年被成功克隆，1999年Schummer等和Wang等通过cDNA微阵列分析才发现HE4基因与卵巢癌有关，展开更多
AIM： To identify alterations in genes and molecular functional pathways in esophageal cancer in a high incidence region of India where there is a widespread use of tobacco and betel quid with fermented areca nuts. ME...AIM： To identify alterations in genes and molecular functional pathways in esophageal cancer in a high incidence region of India where there is a widespread use of tobacco and betel quid with fermented areca nuts. METHODS： Total RNA was isolated from tumor and matched normal tissue of 16 patients with esophageal squamous cell carcinoma. Pooled tumor tissue RNA was labeled with Cy3-dUTP and pooled normal tissue RNA was labeled with Cy5-dUTP by direct labeling method. The labeled probes were hybridized with human 10K cDNA chip and expression profiles were analyzed by Genespring GX V 7.3 （Silicon Genetics）. RESULTS： Nine hundred twenty three genes were differentially expressed. Of these, 611 genes were upregulated and 312 genes were downregulated. Using stringent criteria （P ≤ 0.05 and ≥ 1.5 fold change）, 127 differentially expressed genes （87 upregulated and 40 downregulated） were identified in tumor tissue. On the basis of Gene Ontology, four different molecular functional pathways （HAPK pathway, G-protein coupled receptor family, ion transport activity, and serine or threonine kinase activity）were most significantly upregulated and six different molecular functional pathways （structural constituent of ribosome, endopeptidase inhibitor activity, structural constituent of cytoskeleton, antioxidant activity, acyl group transferase activity, eukaryotic translation elongation factor activity）were most significantly downregulated. CONCLUSION： Several genes that showed alterations in our study have also been reported from a high incidence area of esophageal cancer in China. This indicates that molecular profiles of esophageal cancer in these two different geographic locations are highly consistent.展开更多
基金the Ministry of Science and Technology of China(JY03-A-10,2002AA2Z1003 , 2003CB114303) Science and Technology Committee of Shanghai Municipality(03DJ14016).
文摘We used cDNA microarrays containing ～9,000 unigenes to identify 486 salt responsive expressed sequence tags (ESTs) (representing ～450 unigenes) in shoots of the highly salt-tolerant rice variety, Nona Bokra (Oryza sativa L. Ssp.Indica pv. Nona). Some of the genes identified in this study had previously been associated with salt stress. Howeverthe majority were novel, indicating that there is a great number of genes that are induced by salt exposure. Analysis of the salt stress expression profile data of Nona provided clues regarding some putative cellular and molecular processes that are undertaken by this tolerant rice variety in response to salt stress. Namely, we found that multiple transcription factors were induced during the initial salt response of shoots. Many genes whose encoded proteins are implicated in detoxification, protectant and transport were rapidly induced. Genes supporting photosynthesis were repressed and those supporting carbohydrate metabolism were altered. Commonality among the genes induced by salt exposure with those induced during senescence and biotic stress responses suggests that there are shared signaling pathways among these processes. We further compared the transcriptome changes of the salt-sensitive cultivar, IR28, with that of Nona rice. Many genes that are salt responsive in Nona were found to be differentially regulated in IR28. This study identified a large number of candidate functional genes that appear to be involved in salt tolerance and further examination of these genes may enable the molecular basis of salt tolerance to be elucidated.
文摘【目的】探讨菜心抗虫性的分子机制,为抗虫品种选育和抗虫基因挖掘提供理论支持和中间材料。【方法】选用抗小菜蛾品种65和感小菜蛾品种69菜心为亲本构建F2作图群体,利用cDNA-AFLP标记构建转录图谱、分析差异基因表达,筛选和克隆抗虫相关转录衍生片段。【结果】14对引物组合共扩增出330条扩增带,其中238条为多态性带,多态性条带比例为56.00%~100.00%,平均为73.86%,平均每对引物扩增24条带,多态性条带17个。构建的cDNA-AFLP转录图谱由5个连锁群组成,193个标记未能定位在连锁群上,42个位点定位在5个连锁群上,总长度为575.869 c M,平均图距为115.1738 c M。每个连锁群上的标记数为2~23个,平均为8个。5个连锁群的LOD值变化范围为9.923~12.71,平均为13.207。A1连锁群上的分子标记分布不均匀,出现成簇分布现象和出现了5个较大的断裂。共有18个偏分离位点,偏分离比例为42.86%。在330个位点中,异常分离位点103个,占总位点数的34.24%。对克隆测序成功的10个转录衍生片段分析表明,1个TDF5-1-1与防御相关基因GW775571.1具有高度的序列同源性,8个TDF与大白菜花蕾败育相关基因GR308173.1高度同源,1个与功能未知基因EX106546.1高度的序列同源。【结论】cDNA-AFLP标记能可初步从转录组水平上揭示抗虫菜心群体的连锁遗传规律及抗虫分子机制,其抗性相关基因可进一步用于小菜蛾抗性育种。
文摘番茄果实富含的次生物质严重干扰其总RNA的提取,本试验采用4种方法对番茄果实总RNA进行提取,筛选出一种适合于番茄后续的cDNA-扩增片段长度多态性(amplified fragment length polymorphism,AFLP)分析的高质量RNA提取方法,该方法排除了蛋白、多糖、以及多酚等次生物质的干扰,总RNA得率高、完整性好、纯度高。通过cDNA-AFLP分析方法的检测,得到了一组条带清晰和多态性丰富的图谱。
文摘目前肿瘤癌原125（CA125）检测被广泛应用于临床卵巢癌的诊断，但由于子宫内膜异位症、盆腔炎症等也会导致CA125升高，因此，需要一种敏感性和特异性较强的标志物来提高卵巢癌的诊断水平。人附睾蛋白4（human epididymis protein 4，HFA）基因最早是从人附睾上皮细胞中发现，1991年被成功克隆，1999年Schummer等和Wang等通过cDNA微阵列分析才发现HE4基因与卵巢癌有关，
基金Supported by Non Communicable Disease Division,Indian Council of Medical Research
文摘AIM： To identify alterations in genes and molecular functional pathways in esophageal cancer in a high incidence region of India where there is a widespread use of tobacco and betel quid with fermented areca nuts. METHODS： Total RNA was isolated from tumor and matched normal tissue of 16 patients with esophageal squamous cell carcinoma. Pooled tumor tissue RNA was labeled with Cy3-dUTP and pooled normal tissue RNA was labeled with Cy5-dUTP by direct labeling method. The labeled probes were hybridized with human 10K cDNA chip and expression profiles were analyzed by Genespring GX V 7.3 （Silicon Genetics）. RESULTS： Nine hundred twenty three genes were differentially expressed. Of these, 611 genes were upregulated and 312 genes were downregulated. Using stringent criteria （P ≤ 0.05 and ≥ 1.5 fold change）, 127 differentially expressed genes （87 upregulated and 40 downregulated） were identified in tumor tissue. On the basis of Gene Ontology, four different molecular functional pathways （HAPK pathway, G-protein coupled receptor family, ion transport activity, and serine or threonine kinase activity）were most significantly upregulated and six different molecular functional pathways （structural constituent of ribosome, endopeptidase inhibitor activity, structural constituent of cytoskeleton, antioxidant activity, acyl group transferase activity, eukaryotic translation elongation factor activity）were most significantly downregulated. CONCLUSION： Several genes that showed alterations in our study have also been reported from a high incidence area of esophageal cancer in China. This indicates that molecular profiles of esophageal cancer in these two different geographic locations are highly consistent.
