目的:得到冬凌草高通量转录组数据,为挖掘冬凌草二萜类生物合成途径关键基因,研究冬凌草ESTSSR分子标记提供数据来源。方法:利用Illumina Hi Seq 4000高通量测序技术,采用Trinity的方法从头组装、序列拼接和去冗余处理;基于BLAST完成Uni...目的:得到冬凌草高通量转录组数据,为挖掘冬凌草二萜类生物合成途径关键基因,研究冬凌草ESTSSR分子标记提供数据来源。方法:利用Illumina Hi Seq 4000高通量测序技术,采用Trinity的方法从头组装、序列拼接和去冗余处理;基于BLAST完成Unigene编码蛋白NR功能注释、GO分类、KEGG代谢通路注释和分类、功能基因挖掘,SSR分子标记挖掘。结果:获得冬凌草叶与茎两种不同组织共12 GB转录组数据,得到3 7961个Unigene基因,平均长度1063 bp;得到冬凌草叶和茎转录组有60条unigenes参与萜类化合物骨架合成、6条unigene参与各种萜类化合物合成,有26条unigene参与二萜化合物合成途径,叶与茎两种组织中有差异表达的基因4565个,其中在茎中表达较高的为1668个,在叶中表达较高的有2697个,与二萜类化合物合成相关的差异表达基因有15个。结论:本研究为冬凌草二萜化合物生物合成途径中关键基因的挖掘和分子育种提供了数据信息,为深入研究冬凌草中冬凌草甲素等有效成分的生物合成途径及其调控机制提供基础。展开更多
Objective To construct a morphine tolerance model in primarily cultured striatal neurons, and screen the differentially expressed genes in this model using suppression subtractive hybridization （SSH）. Methods Sbtra...Objective To construct a morphine tolerance model in primarily cultured striatal neurons, and screen the differentially expressed genes in this model using suppression subtractive hybridization （SSH）. Methods Sbtracted cDNA libraries were constructed using SSH from normal primarily cultured striatal neurons and long-term morphine treated striatal neurons （10^-5 mol/L for 72 hours）. To check reliability of the cell culture model, RT-PCR was performed to detect the cAMP-responsive element-binding protein （CREB） mRNA expression. The subtracted clones were prescreened by PCR. The clones containing inserted fragments from forward libraries were sequenced and submitted to GenBank for homology analysis. And the expression levels of genes of interest were confirmed by RT-PCR. Results CREB mRNA expression showed a significant increase in morphine treated striatal neurons （62.85± 1.98） compared with normal striatal neurons （28.43 ± 1.46, P〈0.01）. Thirty-six clones containing inserted fragments were randomly chosen for sequence analysis. And the 36 clones showed homology with 19 known genes and 2 novel genes. The expression of 2 novel genes, mitochondrial carrier homolog 1 （Mtchl ; 96.81±2.04 vs. 44.20±1.31, P〈0.01） and thyrnoma viral proto-oncogene 1 （Akt1 ; 122.10±2.17 vs 50.11±2.01, P〈0.01）, showed a significant increase in morphine-treated striatal neurons compared with normal striatal neurons. Conclusions A reliable differential cDNA library of striatal neurons treated with long-term morphine is constructed. Mtchl and Aktl might be the candidate genes for the development of morphine tolerance.展开更多
Objectives To explore the intrinsic factors related to the pathogenesis of acute arterial thrombosis （AAT） and to elucidate the patho- genesis of AAT on the basis of differentially expressed genes. Methods Patients ...Objectives To explore the intrinsic factors related to the pathogenesis of acute arterial thrombosis （AAT） and to elucidate the patho- genesis of AAT on the basis of differentially expressed genes. Methods Patients with acute myocardial infarction （AMI）, stable angina （SA） and healthy controls （n = 20 per group） were recruited, and the whole human genome microarray analysis was performed to detect the differentially expressed genes among these subjects. Results Patients with AMI had disease-specific gene expression pattern. Biological functional analysis showed the function of T cells was significantly reduced, the mitochondrial metabolism significantly decreased, the ion metabolism was abnormal, the cell apoptosis and inflammatory reaction increased, the phagocytosis elevated, the neutrophil-mediated immunity increased and the post-traumatic repair of cells and tissues increased in AMI patients. The biological function in SA group and healthy controis remained stable and was comparable. Conclusions The reduced function ofT cell gene models in AAT showed the dysfunction of the immune system. The pathogenesis of AAT may be related to the inflammatory reaction after arterial intima infection caused by potential pathogenic microorganisms.展开更多
Suppresssion subtractive hybridization (SSH) was preformed to compare gene expression profiles of PIH patients and normal pregnancy placentas. The subtractive cDNA library of PIH placenta was set up and screedned. Dif...Suppresssion subtractive hybridization (SSH) was preformed to compare gene expression profiles of PIH patients and normal pregnancy placentas. The subtractive cDNA library of PIH placenta was set up and screedned. Differential cDNAs were cloned, and sequenced by T 7 primer methodology. One hundred and three differential cDNAs were isolated by SSH. Sequencing and BLAST analysis showed 90 inserts shared more than 95% homolog with sequences in the GenBank/EMBL database. We identified 36 putative genes including pregnancy-specific glycoprotein gene (BC005924), serine protease inhibitor gene(BC012868), VEGFR-1 gene(AF063657, etc.展开更多
Objective: To analyze the differential expression genes (DEGs) between Barrett’s esophagus (BE) and normal esophagus mucosa and explore the target genes related to the development and progression of BE. Methods: The ...Objective: To analyze the differential expression genes (DEGs) between Barrett’s esophagus (BE) and normal esophagus mucosa and explore the target genes related to the development and progression of BE. Methods: The total RNAs of matched BE and normal esophagus mucosa of BE patients were isolated using one step Trizol method. Matched RNAs were qualified using 10 g/L agarose gel electrophoresis. cRNAs were synthesized, fluorescence labeled and purified after total RNAs were purified. The RNAs of BE and normal esophagus mucosa were hybridized with Agilent oligomicroarray (30 968 probes). The fluorescence intensity features were detected by Agilent scanner and quantified by feature extraction software. Results: (1) The total RNA, reverse transcription product and fluorescence labeled cRNA were all of high quality; (2) There were 142 up-regulated genes and 284 down-regulated genes among 2-fold DEGs. Conclusion: Microarray-based studies are feasible in endoscopically obtained tissues. Many BE-associated genes are screened by the high-throughput gene chip. The development and progression of BE is a complicated process involving multiple genes and multiple procedures, and functional study of these genes may help to identify the key genes or pathways involved in the pathogenesis and development of BE.展开更多
基金Supported by the National Natural Science Foundation of China (81070961,30770676,and 30870932)the Natural Science Foundation of Shandong Province (ZR2009DZ004)the Science and Technology Bureau Foundation of Shandong Province (2006GG2202037)
文摘Objective To construct a morphine tolerance model in primarily cultured striatal neurons, and screen the differentially expressed genes in this model using suppression subtractive hybridization （SSH）. Methods Sbtracted cDNA libraries were constructed using SSH from normal primarily cultured striatal neurons and long-term morphine treated striatal neurons （10^-5 mol/L for 72 hours）. To check reliability of the cell culture model, RT-PCR was performed to detect the cAMP-responsive element-binding protein （CREB） mRNA expression. The subtracted clones were prescreened by PCR. The clones containing inserted fragments from forward libraries were sequenced and submitted to GenBank for homology analysis. And the expression levels of genes of interest were confirmed by RT-PCR. Results CREB mRNA expression showed a significant increase in morphine treated striatal neurons （62.85± 1.98） compared with normal striatal neurons （28.43 ± 1.46, P〈0.01）. Thirty-six clones containing inserted fragments were randomly chosen for sequence analysis. And the 36 clones showed homology with 19 known genes and 2 novel genes. The expression of 2 novel genes, mitochondrial carrier homolog 1 （Mtchl ; 96.81±2.04 vs. 44.20±1.31, P〈0.01） and thyrnoma viral proto-oncogene 1 （Akt1 ; 122.10±2.17 vs 50.11±2.01, P〈0.01）, showed a significant increase in morphine-treated striatal neurons compared with normal striatal neurons. Conclusions A reliable differential cDNA library of striatal neurons treated with long-term morphine is constructed. Mtchl and Aktl might be the candidate genes for the development of morphine tolerance.
文摘Objectives To explore the intrinsic factors related to the pathogenesis of acute arterial thrombosis （AAT） and to elucidate the patho- genesis of AAT on the basis of differentially expressed genes. Methods Patients with acute myocardial infarction （AMI）, stable angina （SA） and healthy controls （n = 20 per group） were recruited, and the whole human genome microarray analysis was performed to detect the differentially expressed genes among these subjects. Results Patients with AMI had disease-specific gene expression pattern. Biological functional analysis showed the function of T cells was significantly reduced, the mitochondrial metabolism significantly decreased, the ion metabolism was abnormal, the cell apoptosis and inflammatory reaction increased, the phagocytosis elevated, the neutrophil-mediated immunity increased and the post-traumatic repair of cells and tissues increased in AMI patients. The biological function in SA group and healthy controis remained stable and was comparable. Conclusions The reduced function ofT cell gene models in AAT showed the dysfunction of the immune system. The pathogenesis of AAT may be related to the inflammatory reaction after arterial intima infection caused by potential pathogenic microorganisms.
基金Supported in part by the National Natural Science Foundation of China (30070787)
文摘Suppresssion subtractive hybridization (SSH) was preformed to compare gene expression profiles of PIH patients and normal pregnancy placentas. The subtractive cDNA library of PIH placenta was set up and screedned. Differential cDNAs were cloned, and sequenced by T 7 primer methodology. One hundred and three differential cDNAs were isolated by SSH. Sequencing and BLAST analysis showed 90 inserts shared more than 95% homolog with sequences in the GenBank/EMBL database. We identified 36 putative genes including pregnancy-specific glycoprotein gene (BC005924), serine protease inhibitor gene(BC012868), VEGFR-1 gene(AF063657, etc.
文摘Objective: To analyze the differential expression genes (DEGs) between Barrett’s esophagus (BE) and normal esophagus mucosa and explore the target genes related to the development and progression of BE. Methods: The total RNAs of matched BE and normal esophagus mucosa of BE patients were isolated using one step Trizol method. Matched RNAs were qualified using 10 g/L agarose gel electrophoresis. cRNAs were synthesized, fluorescence labeled and purified after total RNAs were purified. The RNAs of BE and normal esophagus mucosa were hybridized with Agilent oligomicroarray (30 968 probes). The fluorescence intensity features were detected by Agilent scanner and quantified by feature extraction software. Results: (1) The total RNA, reverse transcription product and fluorescence labeled cRNA were all of high quality; (2) There were 142 up-regulated genes and 284 down-regulated genes among 2-fold DEGs. Conclusion: Microarray-based studies are feasible in endoscopically obtained tissues. Many BE-associated genes are screened by the high-throughput gene chip. The development and progression of BE is a complicated process involving multiple genes and multiple procedures, and functional study of these genes may help to identify the key genes or pathways involved in the pathogenesis and development of BE.
