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Identification of Differentially Expressed Genes Associated with Cotton Fiber Development in a Chromosomal Substitution Line(CS-B22sh) 被引量:4
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作者 SOLIMAN Khairy M BOLTON James J SAHA Sukumar JENKINS Johnie N 《棉花学报》 CSCD 北大核心 2008年第S1期36-,共1页
One of the impediments in the genetic improvement of cotton fiber is the paucity of information about genes associated with fiber development.Availability of chromosome arm substitution line CS-
关键词 CS-B22sh Identification of Differentially expressed genes Associated with Cotton Fiber Development in a Chromosomal Substitution Line LINE
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Analysis on differentially expressed genes in watermelon rind color based on RNA–Seq
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作者 杨侃侃 梁志怀 吴才君 《Journal of Central South University》 SCIE EI CAS CSCD 2016年第11期2818-2826,共9页
In order to screen the genes controlling watermelon rind color and luster, the experiment was carried out with yellow watermelon skin mutants as tester and green wild type watermelon as control, and transcriptome sequ... In order to screen the genes controlling watermelon rind color and luster, the experiment was carried out with yellow watermelon skin mutants as tester and green wild type watermelon as control, and transcriptome sequencing and bioinformatics analysis were done. The results show that 34.27 Gb clean data were got by transcriptome sequencing. There are 261 differentially expressed genes among Y_1_vs_G_1, Y_2_vs_G_2 and Y_3_vs_G_3. The pathways contenting most differentially expressed genes are plant hormone signal transduction pathway, phenylpropanoid biosynthesis pathway, photosynthesis pathway, starch and sucrose metabolism pathway. 9-cis-epoxycarotenoid dioxygenase(Cla002942), alcohol dehydrogenase(Cla004992), photosystem Ⅰ reaction center subunit Ⅲ, chloroplastic(precursor)(Cla009181), long-chain acyl coenzyme A synthetase(Cla017341), threonine dehydratase biosynthetic(Cla018352) candidates genes were screened out. 展开更多
关键词 WATERMELON rind color transcriptome sequencing differentially expressed genes
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Identification of differentially expressed genes in omental adipose tissues of obese patients by suppression subtractive hybridization 被引量:3
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作者 Qiu, j. Ni, Y. H. Gong, H. X. Fei, L. Pan, X. Q. Guo, M. Chen, R. H. Guo, X. R. 《南京医科大学学报(自然科学版)》 CAS CSCD 北大核心 2007年第10期1122-1122,共1页
关键词 基因表达 网膜 脂肪组织 抑制减法杂交 肥胖 鉴别
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Identification of differentially expressed genes in omental adipose tissues of obese patients by suppression subtractive hybridization 被引量:3
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作者 Qiu, J. Ni, Y. H. +5 位作者 Gong, H. X. Fei, L. Pan, X. Q. Guo, M. Chen, R. H. Guo, X. R. 《南京医科大学学报(自然科学版)》 CAS CSCD 北大核心 2007年第5期427-427,共1页
关键词 肥胖症 脂肪组织 削减杂交 差异表达基因 葡萄糖转运泵
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Identification and analysis of differentially expressed genes involved in dark-induced photoperiod response and senescence of soybean leaves by suppression subtractive hybridization
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作者 ZHAO Lin1,LUO Qiu-lan1,HAN Ying-peng1,YANG Chun-liang2,LI Wen-bin1(1.Soybean Research Institute(Key Laboratory of Soybean Biology of Chinese Education Ministry),Northeast Agriculture University,Haerbin,China 150030 2.The Basic MedicalScience College,Harbin Medical University,Haerbin,China 150081) 《湖南农业大学学报(自然科学版)》 CAS CSCD 北大核心 2007年第S1期222-,共1页
A cDNA subtractive library enriched for dark-induced up-regulated ESTs was constructed by suppression subtractive hybridization(SSH) from leaf tissues of soybean cultivar DongNong L13 treated with short-day(8-h light/... A cDNA subtractive library enriched for dark-induced up-regulated ESTs was constructed by suppression subtractive hybridization(SSH) from leaf tissues of soybean cultivar DongNong L13 treated with short-day(8-h light/16-h dark) and long-day(16-h light/8-h dark) conditions.A total of 148 clones were sequenced,representing 76 unique ESTs which corresponded to about 20% of 738 clones from the cDNA library and showed a significant up-regulation of at least three fold verified by dot blot hybridization.The putative functions of ESTs were predicted by Blastn and Blastx.The 43 differentially expressed genes identified by subtractions were classified according to their putative functions generated by Blast analysis.Genetic functional analysis indicated that putative proteins encoded by these genes were related to diverse functions during organism development,which include biological regulation pathways such as transcription,signal transduction and programmed cell death,protein,nucleic acid and carbohydrate macromolecule degradation,the cell wall modification,primary and secondary metabolism and stress response.Two soybean transcription factors enhanced in SD conditions,GAMYB-binding protein and DNA binding protein RAV cDNAs that may be involved in SD soybean photoperiod response,had been isolated using 5'-and 3'-rapid amplification of cDNA ends(RACE)(Genbank Accession numbers DQ112540 and DQ147914). 展开更多
关键词 cDNA Identification and analysis of differentially expressed genes involved in dark-induced photoperiod response and senescence of soybean leaves by suppression subtractive hybridization ESTs
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Different Techniques of Screening Differentially Expressed Genes and Their Applying in Tea Plant
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作者 Xun-Lei Wang Jian-Liang Lu Yue-Rong Liang 《茶叶》 2013年第4期223-230,共8页
The genome of common varieties of tea plant,with estimated size 4000 Mb,is not clearly yet.In order to understand the gene expression difference during certain biological process,we can use transcriptome which reflect... The genome of common varieties of tea plant,with estimated size 4000 Mb,is not clearly yet.In order to understand the gene expression difference during certain biological process,we can use transcriptome which reflects the gene expression level and is more closely related to current status instead of genome.This paper reviews several widely used techniques to screen differentially expressed genes in transcriptome level by comparing the characteristics of the methods and their application in tea plant research. 展开更多
关键词 差异表达基因 茶树 筛选 技术 基因表达差异 转录水平 生物过程 基因组
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Genome-wide identification of Gossypium INDETERMINATE DOMAIN genes and their expression profiles in ovule development and abiotic stress responses 被引量:7
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作者 ALI Faiza QANMBER Ghulam +5 位作者 LI Yonghui MA Shuya LU Lili YANG Zuoren WANG Zhi LI Fuguang 《Journal of Cotton Research》 2019年第1期1-16,共16页
Background:INDETERMINATE DOMAIN(IDD)transcription factors form one of the largest and most conserved gene families in plant kingdom and play important roles in various processes of plant growth and development,such as... Background:INDETERMINATE DOMAIN(IDD)transcription factors form one of the largest and most conserved gene families in plant kingdom and play important roles in various processes of plant growth and development,such as flower induction in term of flowering control.Till date,systematic and functional analysis of IDD genes remained infancy in cotton.Results:In this study,we identified total of 162 IDD genes from eight different plant species including 65 IDD genes in Gossypium hirsutum.Phylogenetic analysis divided IDDs genes into seven well distinct groups.The gene structures and conserved motifs of GhIDD genes depicted highly conserved exon-intron and protein motif distribution patterns.Gene duplication analysis revealed that among 142 orthologous gene pairs,54 pairs have been derived by segmental duplication events and four pairs by tandem duplication events.Further,Ka/Ks values of most of orthologous/paralogous gene pairs were less than one suggested the purifying selection pressure during evolution.Spatiotemporal expression pattern by qRT-PCR revealed that most of the investigated GhIDD genes showed higher transcript levels in ovule of seven days post anthesis,and upregulated response under the treatments of multiple abiotic stresses.Conclusions:Evolutionary analysis revealed that IDD gene family was highly conserved in plant during the rapid phase of evolution.Whole genome duplication,segmental as well as tandem duplication significantly contributed to the expansion of IDD gene family in upland cotton.Some distinct genes evolved into special subfamily and indicated potential role in the allotetraploidy Gossypium hisutum evolution and development High transcript levels of GhIDD genes in ovules illustrated their potential roles in seed and fiber development Further,upregulated responses of GhIDD genes under the treatments of various abiotic stresses suggested them as important genetic regulators to improve stress resistance in cotton breeding. 展开更多
关键词 UPLAND cotton IDD transcription factor gene DUPLICATION COLLINEARITY Spatiotemporal expression Abiotic stresses
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Genome-wide identification and expression profiling of photosystem II(PsbX)gene family in upland cotton(Gossypium hirsutum L.)
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作者 RAZA Irum PARVEEN Abida +4 位作者 AHMAD Adeel HU Daowu PAN Zhaoe ALI Imran DU Xiongming 《Journal of Cotton Research》 CAS 2024年第1期1-14,共14页
Background Photosystem II(PSII)constitutes an intricate assembly of protein pigments,featuring extrinsic and intrinsic polypeptides within the photosynthetic membrane.The low-molecular-weight transmembrane protein Psb... Background Photosystem II(PSII)constitutes an intricate assembly of protein pigments,featuring extrinsic and intrinsic polypeptides within the photosynthetic membrane.The low-molecular-weight transmembrane protein PsbX has been identified in PSII,which is associated with the oxygen-evolving complex.The expression of PsbX gene protein is regulated by light.PsbX’s central role involves the regulation of PSII,facilitating the binding of quinone molecules to the Qb(PsbA)site,and it additionally plays a crucial role in optimizing the efficiency of photosynthesis.Despite these insights,a comprehensive understanding of the PsbX gene’s functions has remained elusive.Results In this study,we identified ten PsbX genes in Gossypium hirsutum L.The phylogenetic analysis results showed that 40 genes from nine species were classified into one clade.The resulting sequence logos exhibited substantial conservation across the N and C terminals at multiple sites among all Gossypium species.Furthermore,the ortholo-gous/paralogous,Ka/Ks ratio revealed that cotton PsbX genes subjected to positive as well as purifying selection pressure might lead to limited divergence,which resulted in the whole genome and segmental duplication.The expression patterns of GhPsbX genes exhibited variations across specific tissues,as indicated by the analysis.Moreover,the expression of GhPsbX genes could potentially be regulated in response to salt,intense light,and drought stresses.Therefore,GhPsbX genes may play a significant role in the modulation of photosynthesis under adverse abiotic conditions.Conclusion We examined the structure and function of PsbX gene family very first by using comparative genom-ics and systems biology approaches in cotton.It seems that PsbX gene family plays a vital role during the growth and development of cotton under stress conditions.Collectively,the results of this study provide basic information to unveil the molecular and physiological function of PsbX genes of cotton plants. 展开更多
关键词 PHOTOSYSTEM PHYLOgeneTIC SYNTENY RNA seq gene expression Orthologous
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Expression Profiling Identifies Candidate Genes for Fiber Yield and Quality 被引量:3
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作者 LLEWELLYN D J MACHADO A +1 位作者 AI-GHAZI Y DENNIS E S 《棉花学报》 CSCD 北大核心 2008年第S1期9-,共1页
Gene expression profiling at early stages(0~2 DPA) of fiber development in Gossypium hirsutum identified a number of transcription factors which were down regulated in fiberless mutants relative to wild type controls... Gene expression profiling at early stages(0~2 DPA) of fiber development in Gossypium hirsutum identified a number of transcription factors which were down regulated in fiberless mutants relative to wild type controls and which could play a role in controlling early fiber development.Chief among these was GhMYB25,a Mixta-like MYB gene.Transgenic GhMYB25-silenced cotton 展开更多
关键词 gene ROOT expression Profiling Identifies Candidate genes for Fiber Yield and Quality
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Genome-wide identification and expression analysis of Gossypium RING-H2 finger E3 ligase genes revealed their roles in fiber development,and phytohormone and abiotic stress responses 被引量:6
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作者 QANMBER Ghulam YU Daoqian +5 位作者 LI Jie WANG Lingling MA Shuya LU Lili YANG Zuoren LI Fuguang 《Journal of Cotton Research》 2018年第1期3-19,共17页
Background: RING H2 finger E3 ligase (RH2FE3) genes encode cysteine rich proteins that mediate E3 ubiquitin ligase activity and degrade target substrates. The roles of these genes in plant responses to phytohormone... Background: RING H2 finger E3 ligase (RH2FE3) genes encode cysteine rich proteins that mediate E3 ubiquitin ligase activity and degrade target substrates. The roles of these genes in plant responses to phytohormones and abiotic stresses are well documented in various species, but their roles in cotton fiber development are poorly understood. To date, genome wide identification and expression analyses of Gossypium hirsutum RH2FE3 genes have not been reported. Methods: We performed computational identification, structural and phylogenetic analyses, chromosomal distribution analysis and estimated KJKs values of G hirsutum RH2FE3 genes. Orthologous and paralogous gene pairs were identified by all versus all BLASTP searches. We predicted cis regulatory elements and analyzed microarray data sets to generate heatmaps at different development stages. Tissue specific expression in cotton fiber, and hormonal and abiotic stress responses were determined by quantitative real time polymerase chain reaction (qRT PCR) analysis. Results: We investigated 140 G hirsutum, 80 G. orboreum, and evolutionary mechanisms and compared them with orthologs 89 G. roimondii putative RH2FB genes and their in Arobidopsis and rice. A domain based analysis of the G hirsutum RH2FE3 genes predicted conserved signature motifs and gene structures. Chromosomal localization showed the genes were distributed across all G hirsutum chromosomes, and 60 duplication events (4 tandem and 56 segmental duplications) and 98 orthologs were detected, cis elements were detected in the promoter regions of G hirsutum RH2FE3 genes. Microarray data and qRT PCR analyses showed that G hirsutum RH2FE3 genes were strongly correlated with cotton fiber development. Additionally, almost all the (brassinolide, gibberellic acid (GA), indole 3-acetic acid drought, and salt). dentified genes were up regulated in response to phytohormones (IAA), and salicylic acid (SA)) and abiotic stresses (cold, heat, Conclusions: The genome wide identification, comprehensive analysis, and characterization of conserved domains and gene structures, as well as phylogenetic analysis, cis element prediction, and expression profile analysis of G hirsutum RH2FE3 genes and their roles in cotton fiber development and responses to plant hormones and abiotic stresses are reported here for the first time. Our findings will contribute to the genome wide analysis of putative RH2FE3 genes in other species and lay a foundation for future physiological and functional research on G hirsutum RH2FE3 genes. 展开更多
关键词 Gossypium hirsutum Upland cotton RING H2 finger E3 ligase Phylogenetic analysis cis elements gene duplication expression profile analysis
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Characterization of the 19 Novel Cotton FLA Genes and Their Expression Profiling in Fiber Development and in Response to Phytohormones and Salt Stress 被引量:12
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作者 HUANG Geng-qing,XU Wen-liang,GONG Si-ying,WANG Xiu-lan,LI Xue-bao(College of Life Sciences,Huazhong Normal University,Wuhan 430079,China) 《棉花学报》 CSCD 北大核心 2008年第S1期55-,共1页
Fasciclin-like arabinogalactan proteins(FLAs),a subclass of arabinogalactan proteins(AGPs),are usually involved in cell development in plants.To investigate the expression profiling as well
关键词 FLA Characterization of the 19 Novel Cotton FLA genes and Their expression Profiling in Fiber Development and in Response to Phytohormones and Salt Stress
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Identification, characterization, and expression profiles of the GASA genes in cotton 被引量:2
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作者 QIAO Kaikai MA Changkai +3 位作者 LV Jiaoyan ZHANG Chaojun MA Qifeng FAN Shuli 《Journal of Cotton Research》 2021年第1期50-65,共16页
Background:GASA(Giberellic Acid Stimulated in Arabidopsis)gene family plays a crucial role in the phytohormone signaling pathway,growth and development,and stress responses in plants.Many GASA homologs have been ident... Background:GASA(Giberellic Acid Stimulated in Arabidopsis)gene family plays a crucial role in the phytohormone signaling pathway,growth and development,and stress responses in plants.Many GASA homologs have been identified in various plants.Nevertheless,little is known about these proteins in cotton.Results:In the current study,we identified 19,17,25,33,and 38 GASA genes via genome-wide analyses of Gossypium herbaceum,G.arboreum,G.raimondii,G.barbadense,and G.hirsutum,respectively,and performed comprehensive bioinformatics and expression analyses.According to our results,132 GASA proteins shared similar protein structures and were classified into four groups based on the phylogenetic tree.A synteny analysis suggested that segmental duplication was a key driver in the expansion of the GASA gene family.Meanwhile,the cis-element and protein interaction analyses indicated that GhGASA proteins play a significant role in the hormone responses.Transcriptomic and qRT-PCR(Quantitative real time-polymerase chain reaction)analyses revealed diverse expression profiles of the GhGASA genes in different organs under abiotic stresses,indicating that some GhGASA genes possibly participate in fiber development and abiotic-stress responses.Conclusions:The GASA genes in cotton were systematically identified and analyzed for the first time in this paper,and it suggested that the GASA genes are important to the development and growth of cotton.These results will support future exploration of the functions of GASA genes in cotton. 展开更多
关键词 GASA gene GENOME-WIDE BIOINFORMATICS expression pattern COTTON
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Expression Profile Analysis of Genes Involved in Brassinosteroid Biosynthesis Pathway in Cotton Fiber Development
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作者 LUO Ming,XIAO Zhong-yi,TAN Kun-ling,HU Ming-yu,LIAO Peng(Key Laboratory of Biotechnology and Crop Quality Improvement,Ministry of Agriculture Biotechnology Research Center,Southwest University,Chongqing 400716,China) 《棉花学报》 CSCD 北大核心 2008年第S1期59-,共1页
Cotton(Gossypium hirsutum L.) is the leading fiber crop and one of the mainstays of the economy in the world.Cotton fibers,as the main product of cotton plants,are unicellular,linear
关键词 DPA expression Profile Analysis of genes Involved in Brassinosteroid Biosynthesis Pathway in Cotton Fiber Development
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Quantification of the expression of chitinolytic enzyme encoding genes ech30, ech42 and nag1 in Trichoderma atroviride P1 under varying growth conditions using a real-time RT-PCR assay
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作者 Jihong Liu Clarke Arne Tronsmo +1 位作者 Nicholas Clarkel Sonja Sletner Klemsdal 《浙江大学学报(农业与生命科学版)》 CAS CSCD 北大核心 2004年第4期428-428,共1页
The quantitative expression and the regulation of chitinase-encoding genes ech30, ech42 and nag1 in Trichoderma atroviride P1 under varying growth conditions were investigated using real-time RT-PCR, principle compone... The quantitative expression and the regulation of chitinase-encoding genes ech30, ech42 and nag1 in Trichoderma atroviride P1 under varying growth conditions were investigated using real-time RT-PCR, principle component and multivariate analyses. Twelve media combinations including 0.1% and 3% glucose as carbon source and no (0 mmol/L), low (10 mmol/L) and high (100 mmol/L) ammonium acetate as nitrogen source combined with or without colloidal chitin at 3 time intervals and 2 replications were applied to current study. The real-time RT-PCR analysis showed that the expression of ech30, ech42 and nag1 was regulated by the interaction of nitrogen, glucose and chitin under different growth conditions. The highest and earliest expressions of ech30 were induced by glucose and nitrogen starvation i.e. 0.1% glucose and 10 mmol/L ammonium acetate in the growth media. This was also the case for ech42 and nag1 but at a relatively low level. In contrast, high (3%) glucose and high (100 mmol/L) ammonium acetate concentrations repressed the expression of all the genes studied. These results were confirmed by principle component and multivariate analyses. The effect of chitin on ech30, ech42 and nag1 expression varied depending on the concentrations of glucose and ammonium acetate. 展开更多
关键词 基因 表达 生长条件 RT-PCR 木霉属 真菌
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Genes Expressing Differences in Upland Cotton Fiber Development Between 12 DPA and 16 DPA
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作者 LU Cai-rui1,YU Shu-xun1,WANG Wu1,2(1.Cotton Research Institute,Chinese Academy of Agricultural Sciences Key Laboratory of Cotton Genetic Improvement,Ministry of Agriculture,Anyang,Henan 455000,China 2.Plant and Environmental Sciences Department,New Mexico State University,Las Cruces,NM 88003 USA) 《棉花学报》 CSCD 北大核心 2008年第S1期70-,共1页
Cotton is the major dominant natural fiber crop on the earth.Although some current cotton genetics and breeding programs had made great progresses in cotton lint yield increases and disease resistance(tolerance),fiber... Cotton is the major dominant natural fiber crop on the earth.Although some current cotton genetics and breeding programs had made great progresses in cotton lint yield increases and disease resistance(tolerance),fiber quality has little improvement.Global understanding genes expression 展开更多
关键词 genes expressing Differences in Upland Cotton Fiber Development Between 12 DPA and 16 DPA
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GEO(Gene Expression Omnibus):高通量基因表达数据库 被引量:9
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作者 刘华 马文丽 郑文岭 《中国生物化学与分子生物学报》 CAS CSCD 北大核心 2007年第3期236-244,共9页
GEO(Gene Expression Omnibus)数据库包括高通量实验数据的广泛分类,有单通道和双通道以微阵列为基础的对mRNA丰度的测定;基因组DNA和蛋白质分子的实验数据;其中包括来自以非阵列为基础的高通量功能基因组学和蛋白质组学技术的数据也被... GEO(Gene Expression Omnibus)数据库包括高通量实验数据的广泛分类,有单通道和双通道以微阵列为基础的对mRNA丰度的测定;基因组DNA和蛋白质分子的实验数据;其中包括来自以非阵列为基础的高通量功能基因组学和蛋白质组学技术的数据也被存档,例如基因表达系列分析(serial analysis of gene expression,SAGE)和蛋白质鉴定技术.迄今为止,GEO数据库包含的数据含概10000个杂交实验和来自30种不同生物体的SAGE库.本文概述了GEO数据库的查询和浏览,数据下载和格式,数据分析,贮存与更新,并着重分析GEO数据浏览器中控制词汇的使用,阐述了GEO数据库的数据挖掘以及GEO在分子生物学领域中的应用前景.GEO可由此公众网址直接登陆http://www.ncbi.nlm.nih.gov/projects/geo/. 展开更多
关键词 基因表达 数据库 控制词汇 数据挖掘
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大鼠Neurogenesin-1基因真核表达载体的构建及在cos-7细胞中的表达 被引量:2
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作者 高维陆 尹宗生 +1 位作者 张胜权 张辉 《安徽医科大学学报》 CAS 北大核心 2009年第4期430-433,共4页
目的克隆大鼠海马中Neurogenesin-1(Ng1)基因片段,构建pSecTag2/HygroB-Ng1真核表达载体,并检测其在cos-7细胞中的表达,为进一步研究该基因对脊髓神经干细胞分化的影响提供实验依据。方法在无RNA酶污染的条件下提取出大鼠海马总RNA。利... 目的克隆大鼠海马中Neurogenesin-1(Ng1)基因片段,构建pSecTag2/HygroB-Ng1真核表达载体,并检测其在cos-7细胞中的表达,为进一步研究该基因对脊髓神经干细胞分化的影响提供实验依据。方法在无RNA酶污染的条件下提取出大鼠海马总RNA。利用逆转录聚合酶链反应扩增出Ng1基因片段。将该基因片段连接到真核表达载体pSecTag2/HygroB,聚合酶链反应初步筛选,双酶切鉴定后送测序。将构建成功的重组真核表达载体转染入cos-7细胞,Westernblot鉴定重组Ng1蛋白的表达。结果逆转录聚合酶链反应成功获得大鼠Ng1cDNA。随机挑选10个重组真核表达载体的克隆,聚合酶链反应筛选出阳性克隆2个,经双酶切鉴定、测序及Blast分析鉴定重组质粒构建成功。脂质体介导转染cos-7细胞48h后,Westernblot鉴定重组Ng1蛋白在cos-7细胞中的表达,在46ku处出现阳性条带。结论大鼠海马Ng1基因的真核表达载体pSecTag2/HygroB-Ng1构建成功,转染cos-7细胞后能够表达重组Ng1蛋白。 展开更多
关键词 大鼠 基因表达 遗传载体 Neurogenesin-1
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来源于Alcaligenes A-6的D-氨基酰化酶基因的合成与融合表达
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作者 侯欣彤 董媛 +5 位作者 林瑞东 于梁 任媛媛 李剑光 高朝辉 滕利荣 《高等学校化学学报》 SCIE EI CAS CSCD 北大核心 2014年第8期1670-1674,共5页
将来源于Alcaligenes A-6的D-氨基酰化酶基因用大肠杆菌中的丰沛密码子替换,利用化学和基于聚合酶链反应(PCR)技术的酶促方法进行基因全合成,利用pET-32a构建重组表达载体pET-dan,转化进E.coil BL21(DE3)中进行融合表达.经SDS-PAGE电泳... 将来源于Alcaligenes A-6的D-氨基酰化酶基因用大肠杆菌中的丰沛密码子替换,利用化学和基于聚合酶链反应(PCR)技术的酶促方法进行基因全合成,利用pET-32a构建重组表达载体pET-dan,转化进E.coil BL21(DE3)中进行融合表达.经SDS-PAGE电泳、Western-blot检测和活性测定发现,D-ANase可在大肠杆菌中高效表达,目的蛋白可达到菌体总蛋白的69.2%,密码子优化后基因构建的工程菌发酵活性为96 U/mL,重组蛋白经超声细胞破碎及Ni2+柱亲和层析纯化,比活可达1692.3 U/mg,纯度可达95%以上. 展开更多
关键词 D-氨基酰化酶 产碱杆菌 基因合成 融合表达
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产碱杆菌Alcaligenes sp.P156中龙胆酸1,2-双加氧酶GdoP的研究
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作者 胡春辉 王菲 +1 位作者 郭立忠 于浩 《青岛农业大学学报(自然科学版)》 2019年第3期176-182,共7页
龙胆酸是芳香族化合物微生物降解的核心代谢产物,龙胆酸降解研究对于芳香族化合物的降解研究具有重要意义。龙胆酸1,2-双加氧酶是催化龙胆酸降解的关键酶。本研究利用双加氧酶催化保守结构域在产碱杆菌P156的基因组上通过多序列比对找... 龙胆酸是芳香族化合物微生物降解的核心代谢产物,龙胆酸降解研究对于芳香族化合物的降解研究具有重要意义。龙胆酸1,2-双加氧酶是催化龙胆酸降解的关键酶。本研究利用双加氧酶催化保守结构域在产碱杆菌P156的基因组上通过多序列比对找到一个龙胆酸双加氧酶基因gdoP;利用分子生物学技术对该基因进行异源表达;并利用重组蛋白对酶学性质和动力学参数进行测定。酶学性质研究表明,GdoP催化反应的最适pH为7.5,最适温度为40℃。GdoP是一个Fe2+依赖型双加氧酶,含有Fe2+离子结合结构域。Fe2+离子能够明显促进GdoP酶活,Cu2+、Cd2+离子对GdoP的酶活有明显抑制。以龙胆酸为底物,GdoP的Km值和Vmax值分别为641μmol/L和23.9U/mg。GdoP具有较强的底物特异性,能够催化龙胆酸开环生成3-马来酰丙酮酸,不能催化5-氨基水杨酸、水杨酸和1-羟基-2-萘酸的转化。本研究系统的研究了龙胆酸1,2-双加氧酶GdoP的酶学性质,有助于更好的研究芳香族化合物的微生物降解过程。 展开更多
关键词 龙胆酸1 2-双加氧酶 产碱杆菌 基因表达 酶学性质
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百子莲体细胞发育受体激酶APSERK1(Somatic Embryogenesis Receptor Kinase1)基因全长克隆及表达分析
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作者 张琰 石玉波 《安徽农学通报》 2018年第21期36-40,共5页
根据前期百子莲(Agapanthus praecox ssp.orientalis)转录组测序分析的结果,获得了1个与胚性能力相关的关键基因Somatic Embryogenesis Receptor Kinase1(SERK1)同源性较高的核心片段。采用cDNA末端快速扩增(rapid amplification of cDN... 根据前期百子莲(Agapanthus praecox ssp.orientalis)转录组测序分析的结果,获得了1个与胚性能力相关的关键基因Somatic Embryogenesis Receptor Kinase1(SERK1)同源性较高的核心片段。采用cDNA末端快速扩增(rapid amplification of cDNA ends,RACE)方法得到了百子莲SERK基因cDNA全长序列,命名为ApSERK1。序列分析表明,百子莲ApSERK1基因开放阅读框(ORF)为1800bp,编码1个含有600个氨基酸的蛋白质。氨基酸同源性比对发现,百子莲ApSERK1蛋白序列与水稻、小兰屿蝴蝶兰、铁皮石斛、椰子树和菠萝等植物蛋白序列的相似性可达到90%以上。实时荧光定量qRT-PCR结果表明,ApSERK1基因有一定时空表达差异,在百子莲的种子、小花梗中表达含量最高,且随着外源生长素浓度的增加,胚性愈伤组织的胚性能力呈现下降趋势。推测ApSERK1基因是衡量植物细胞胚性能力的重要指标。 展开更多
关键词 百子莲 体细胞发育受体激酶 基因克隆 表达分析
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