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转入盐地碱蓬谷胱甘肽转移酶和过氧化氢酶基因增强水稻幼苗对低温胁迫的抗性 被引量:30
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作者 赵凤云 王晓云 +1 位作者 赵彦修 张慧 《植物生理与分子生物学学报》 CAS CSCD 北大核心 2006年第2期231-238,共8页
以水稻(OryzasativaL.)品种中花11号成熟种子为材料,利用农杆菌介导法将盐地碱蓬的GST(谷胱甘肽转移酶)单基因和GST+CAT1(catalase1)双基因转入低温敏感水稻品种中花11号,并对T4代转基因水稻幼苗的抗低温特性进行了分析。结果显示,低温... 以水稻(OryzasativaL.)品种中花11号成熟种子为材料,利用农杆菌介导法将盐地碱蓬的GST(谷胱甘肽转移酶)单基因和GST+CAT1(catalase1)双基因转入低温敏感水稻品种中花11号,并对T4代转基因水稻幼苗的抗低温特性进行了分析。结果显示,低温处理后,转基因植株的GST和CAT活性都比未转入这两种基因的对照高;且PSII最大光化学效率也高于非转基因对照;而H2O2和MDA(malondialdehyde)的含量及细胞膜透性则低于对照。说明转基因水稻幼苗GST和GST+CAT1的表达提高了对低温胁迫的抗性。 展开更多
关键词 转基因水稻 低温胁迫 共表达 过氧化氢酶 谷胱甘肽转移酶
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应用硫氧还蛋白促进外源蛋白在大肠杆菌的可溶性表达 被引量:10
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作者 安乃莉 张智清 +4 位作者 王嵩 刘红兵 曾革非 姚立红 陈爱君 《病毒学报》 CAS CSCD 北大核心 1999年第2期130-135,共6页
为了观察硫氧还蛋白(TrxA)促进外源蛋白在大肠杆菌中可溶性表达的作用,我们从质粒pET-32a(+)上克隆了trxA基因,构建了TrxA表达质粒pT-TrxA。将该质粒与其它蛋白基因的表达质粒共同转化Ecoli并... 为了观察硫氧还蛋白(TrxA)促进外源蛋白在大肠杆菌中可溶性表达的作用,我们从质粒pET-32a(+)上克隆了trxA基因,构建了TrxA表达质粒pT-TrxA。将该质粒与其它蛋白基因的表达质粒共同转化Ecoli并同时获得表达。结果表明,共表达TrxA可以明显促进外源蛋白,如甲状旁腺激素相关蛋白(PTHrP)、PTHrP受体(PTHrP-R)和血管内皮生长因子(VEGF)的可溶性表达。说明共表达TrxA有助于外源蛋白的正确折叠,防止包涵体的形成。 展开更多
关键词 硫氧还蛋白 共表达 外源蛋白 大肠杆菌
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Rice Expression Database(RED):An integrated RNA-Seq-derived gene expression database for rice 被引量:16
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作者 Lin Xia Dong Zou +7 位作者 Jian Sang Xingjian Xu Hongyan Yin Mengwei Li Shuangyang Wu Songnian Hu Lili Hao Zhang Zhang 《Journal of Genetics and Genomics》 SCIE CAS CSCD 2017年第5期235-241,共7页
Rice is one of the most important stable food as well as a monocotyledonous model organism for the plant research community.Here,we present RED(Rice Expression Database;http://expression.ic4r.org),an integrated dat... Rice is one of the most important stable food as well as a monocotyledonous model organism for the plant research community.Here,we present RED(Rice Expression Database;http://expression.ic4r.org),an integrated database of rice gene expression profiles derived entirely from RNA-Seq data.RED features a comprehensive collection of 284 high-quality RNA-Seq experiments,integrates a large number of gene expression profiles and covers a wide range of rice growth stages as well as various treatments.Based on massive expression profiles,RED provides a list of housekeeping and tissue-specific genes and dynamically constructs co-expression networks for gene(s) of interest.Besides,it provides user-friendly web interfaces for querying,browsing and visualizing expression profiles of concerned genes.Together,as a core resource in BIG Data Center,RED bears great utility for characterizing the function of rice genes and better understanding important biological processes and mechanisms underlying complex agronomic traits in rice. 展开更多
关键词 Rice expression database expression profiles Housekeeping gene Tissue-specific gene co-expression network
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MicroTom Metabolic Network: Rewiring Tomato Metabolic Regulatory Network throughout the Growth Cycle 被引量:18
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作者 Yan Li Yang Chen +13 位作者 Lu Zhou Shengjie You Heng Deng Ya Chen Saleh Alseekh Yong Yuan Rao Fu Zixin Zhang Dan Su Alisdair R.Fernie Mondher Bouzayen Tao Ma Mingchun Liu Yang Zhang 《Molecular Plant》 SCIE CAS CSCD 2020年第8期1203-1218,共16页
Tomato(Solanum lycopersicum)is a major horticultural crop worldwide and has emerged as a preeminent model for metabolic research.Although many research efforts have focused on the analysis of metabolite differences be... Tomato(Solanum lycopersicum)is a major horticultural crop worldwide and has emerged as a preeminent model for metabolic research.Although many research efforts have focused on the analysis of metabolite differences between varieties and species,the dynamics of metabolic changes during the tomato growth cycle and the regulatory networks that underlie these changes are poorly understood.In this study,we integrated high-resolution spatio-temporal metabolome and transcriptome data to systematically explore the metabolic landscape across 20 major tomato tissues and growth stages.In the resulting MicroTom Metabolic Network,the 540 detected metabolites and their co-expressed genes could be divided into 10 distinct clusters based on their biological functions.Using this dataset,we constructed a global map of the major metabolic changes that occur throughout the tomato growth cycle and dissected the underlying regulatory network.In addition to verifying previously well-established regulatory networks for important metabolites,we identified novel transcription factors that regulate the biosynthesis of important secondary metabolites such as steroidal glycoalkaloids and flavonoids.Our findings provide insights into spatiotemporal changes in tomato metabolism and generate a valuable resource for the study of metabolic regulatory processes in model plants. 展开更多
关键词 TOMATO METABOLOME transcriptome transcription factor co-expression FLAVONOIDS
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De novo assembly of a Chinese soybean genome 被引量:15
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作者 Yanting Shen Jing Liu +7 位作者 Haiying Geng Jixiang Zhang Yucheng Liu Haikuan Zhang Shilai Xing Jianchang Du Shisong Ma Zhixi Tian 《Science China(Life Sciences)》 SCIE CAS CSCD 2018年第8期871-884,共14页
Soybean was domesticated in China and has become one of the most important oilseed crops. Due to bottlenecks in their introduction and dissemination, soybeans from different geographic areas exhibit extensive genetic ... Soybean was domesticated in China and has become one of the most important oilseed crops. Due to bottlenecks in their introduction and dissemination, soybeans from different geographic areas exhibit extensive genetic diversity. Asia is the largest soybean market; therefore, a high-quality soybean reference genome from this area is critical for soybean research and breeding.Here, we report the de novo assembly and sequence analysis of a Chinese soybean genome for "Zhonghuang 13" by a combination of SMRT, Hi-C and optical mapping data. The assembled genome size is 1.025 Gb with a contig N50 of 3.46 Mb and a scaffold N50 of 51.87 Mb. Comparisons between this genome and the previously reported reference genome(cv. Williams82) uncovered more than 250,000 structure variations. A total of 52,051 protein coding genes and 36,429 transposable elements were annotated for this genome, and a gene co-expression network including 39,967 genes was also established. This high quality Chinese soybean genome and its sequence analysis will provide valuable information for soybean improvement in the future. 展开更多
关键词 de novo soybean genome Zhonghuang 13 Gmax_ZH13 structure variation gene co-expression network
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Genome-wide Analysis of MYB Gene Family in Potato Provides Insights into Tissue-specific Regulation of Anthocyanin Biosynthesis 被引量:15
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作者 Yumei Li Jun Liang +5 位作者 Xiangzhi Zeng Han Guo Youwei Luo Philip Kear Shouming Zhang Guangtao Zhu 《Horticultural Plant Journal》 SCIE CSCD 2021年第2期129-141,共13页
The MYB transcription factor is one of the largest gene families in plants,playing an important role in regulating plant growth,development,response to stress,senescence,and especially the anthocyanin biosynthesis.In ... The MYB transcription factor is one of the largest gene families in plants,playing an important role in regulating plant growth,development,response to stress,senescence,and especially the anthocyanin biosynthesis.In this study,A total of 217 MYB genes,including 901R-MYBs,124 R2R3-MYBs,and 3 R1R2R3-MYBs have been identified from the potato genome.The 1R-MYB and R2R3-MYB family members could be divided into 20 and 35 subgroups respectively.Analysis of gene structure and protein motifs revealed that members within the same subgroup presented similar exon/intron and motif organization,further supporting the results of phylogenetic analysis.Potato is an ideal plant to reveal the tissue-specific anthocyanins biosynthesis regulated by MYB,as the anthocyanins could be accumulated in different tissues,showing colorful phenotypes.Five pairs of colored and colorless tissues,stigma,petal,stem,leaf,and tuber flesh,were applied to the transcriptomic analysis.A total of 70 MYB genes were found to be differentially expressed between colored and colorless tissues,and these differentially expressed genes were suspected to regulate the biosynthesis of anthocyanin of different tissues.Co-expression analysis identified numerous potential interactive regulators of anthocyanins biosynthesis,involving 39 MYBs,24 bHLHs,2 WD-repeats,and 29 biosynthesis genes.Genome-wide association study(GWAS)of tuber flesh color revealed amajor signal at the end of Chromosome 10,which was co-localized with reported I gene(StMYB88),controlling tuber peel color.Analyses of DEGs(Differentially Expression Genes)revealed that both StMYB88 and StMYB89 were closely related to regulating anthocyanin biosynthesis of tuber flesh.This work offers a comprehensive overview of the MYB family in potato and will lay a foundation for the functional validation of these genes in the tissue-specific regulation of anthocyanin biosynthesis. 展开更多
关键词 POTATO MYB transcription factor ANTHOCYANIN differentially expressed co-expression tuber flesh
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Construction and Co-expression of Grass Carp Reovirus VP6 Protein and Enhanced Green Fluorescence Protein in the Insect Cells 被引量:13
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作者 Qin FANG Eng Khuan Seng +1 位作者 Wen DAI Lan-lan ZHANG 《中国病毒学》 CSCD 2007年第5期397-404,共8页
Grass carp reovirus(GCRV),a disaster agent to aquatic animals,belongs to Genus Aquareovirus of family Reoviridea.Sequence analysis revealed GCRV genome segment 8(s8) was 1 296 bp nucleotides in length encoding an inne... Grass carp reovirus(GCRV),a disaster agent to aquatic animals,belongs to Genus Aquareovirus of family Reoviridea.Sequence analysis revealed GCRV genome segment 8(s8) was 1 296 bp nucleotides in length encoding an inner capsid protein VP6 of about 43kDa.To obtain in vitro non-fusion expression of a GCRV VP6 protein containing a molecular of fluorescence reporter,the recombinant baculovirus,which contained the GCRVs8 and eGFP(enhanced green fluorescence protein) genes,was constructed by using the Bac-to-Bac insect expression system.In this study,the whole GCRVs8 and eGFP genes,amplified by PCR,were constructed into a pFastBacDual vector under polyhedron(PH) and p10 promoters,respectively.The constructed dual recombinant plasmid(pFbDGCRVs8/eGFP) was transformed into DH10Bac cells to obtain recombinant Bacmid(AcGCRVs8/eGFP) by transposition.Finally,the recombinant bacluovirus(vAcGCRVs8/eGFP) was obtained from transfected Sf9 insect cells.The green fluorescence that was expressed by transfected Sf9 cells was initially observed 3 days post transfection,and gradually enhanced and extended around 5 days culture in P1(Passage1) stock.The stable high level expression of recombinant protein was observed in P2 and subsequent passage budding virus(BV) stock.Additionally,PCR amplification from P1 and amplified P2 BV stock further confirmed the validity of the dual-recombinant baculovirus.Our results provide a foundation for expression and assembly of the GCRV structural protein in vitro. 展开更多
关键词 草鱼呼肠孤病毒 VP6蛋白 增强绿色荧光蛋白 杆状病毒表达系统 共表达
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CecropinB与Xa21基因共表达提高转基因水稻白叶枯病抗性 被引量:10
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作者 于彦春 张光恒 +5 位作者 郭龙彪 曾大力 赵艳 胡兴明 黄大年 钱前 《中国水稻科学》 CAS CSCD 北大核心 2006年第1期105-108,共4页
为了探明CecropinB与Xa21这两种不同抗病途径的基因单独或共表达后的田间抗白叶枯病表现,利用农杆菌转化技术,获得了这两个基因的单独和共表达后的植株。利用PCR等分子鉴定技术,证明这两个基因已经整合到水稻基因组中。抗病性检测结果表... 为了探明CecropinB与Xa21这两种不同抗病途径的基因单独或共表达后的田间抗白叶枯病表现,利用农杆菌转化技术,获得了这两个基因的单独和共表达后的植株。利用PCR等分子鉴定技术,证明这两个基因已经整合到水稻基因组中。抗病性检测结果表明,Xa21和CecropinB均能提高水稻对白叶枯病的抗性,且Xa21的抗病性较CecropinB明显。Xa21和CecropinB基因共表达后的植株的抗病性比单基因的更强,说明将不同抗性机制的基因共表达,可以在育种上利用以提高水稻的白叶枯病抗性。 展开更多
关键词 抗病基因 转基因 白叶枯病 共表达 抗性 水稻
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水稻生物钟相关基因的表达特征鉴定及其功能预测分析 被引量:13
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作者 董昭桐 雷雅琪 +3 位作者 陈淼林 余婧 朱璐 许杰 《基因组学与应用生物学》 CAS CSCD 北大核心 2016年第3期692-705,共14页
水稻作为我国重要农作物,其营养生长和生殖发育过程都受到严格的生物钟的控制,因此,生物钟基因的表达变化也是决定水稻产量和种子质量的主要决定因素之一。为了更全面系统的了解生物钟基因在水稻生长和发育过程中的功能,本研究采用生物... 水稻作为我国重要农作物,其营养生长和生殖发育过程都受到严格的生物钟的控制,因此,生物钟基因的表达变化也是决定水稻产量和种子质量的主要决定因素之一。为了更全面系统的了解生物钟基因在水稻生长和发育过程中的功能,本研究采用生物信息学共表达方法,筛选和鉴定水稻和拟南芥中生物钟基因,并对比分析了这些生物钟基因在单双子叶模式植物中的可能功能。从水稻表达谱公共数据库中筛选与生物钟基因表达密切相关732个基因,并对筛选出的水稻生物钟相关基因进行表达特性、节律性、生物功能预测及其与拟南芥的对比分析,结果表明水稻和拟南芥的生物钟基因可能都具有在生物钟核心振荡器部分功能相对保守的特点。功能预测分析也还表明水稻生物钟基因参与了8类生物学过程,尤其在应激反应和代谢过程的方面具有明显富集性,从而间接佐证了水稻对于外界环境的响应及其代谢过程具有严格时间调控的分子机理。 展开更多
关键词 水稻 生物钟 共表达分析 应激反应 代谢过程
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Identify the signature genes for diagnose of uveal melanoma by weight gene co-expression network analysis 被引量:10
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作者 Kai Shi Zhi-Tong Bing +4 位作者 Gui-Qun Cao Ling Guo Ya-Na Cao Hai-Ou Jiang Mei-Xia Zhang 《International Journal of Ophthalmology(English edition)》 SCIE CAS 2015年第2期269-274,共6页
AIM: To identify and understand the relationship between co-expression pattern and clinic traits in uveal melanoma, weighted gene co-expression network analysis(WGCNA) is applied to investigate the gene expression lev... AIM: To identify and understand the relationship between co-expression pattern and clinic traits in uveal melanoma, weighted gene co-expression network analysis(WGCNA) is applied to investigate the gene expression levels and patient clinic features. Uveal melanoma is the most common primary eye tumor in adults. Although many studies have identified some important genes and pathways that were relevant to progress of uveal melanoma, the relationship between co-expression and clinic traits in systems level of uveal melanoma is unclear yet. We employ WGCNA to investigate the relationship underlying molecular and phenotype in this study.METHODS: Gene expression profile of uveal melanoma and patient clinic traits were collected from the Gene Expression Omnibus(GEO) database. The gene co-expression is calculated by WGCNA that is the R package software. The package is used to analyze the correlation between pairs of expression levels of genes.The function of the genes were annotated by gene ontology(GO).RESULTS: In this study, we identified four co-expression modules significantly correlated with clinictraits. Module blue positively correlated with radiotherapy treatment. Module purple positively correlates with tumor location(sclera) and negatively correlates with patient age. Module red positively correlates with sclera and negatively correlates with thickness of tumor. Module black positively correlates with the largest tumor diameter(LTD). Additionally, we identified the hug gene(top connectivity with other genes) in each module. The hub gene RPS15 A, PTGDS, CD53 and MSI2 might play a vital role in progress of uveal melanoma.CONCLUSION: From WGCNA analysis and hub gene calculation, we identified RPS15 A, PTGDS, CD53 and MSI2 might be target or diagnosis for uveal melanoma. 展开更多
关键词 weighted gene co-expression network analysis microarray data gene ontology
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Glucocorticoids Significantly Influence the Transcriptome of Bone Microvascular Endothelial Cells of Human Femoral Head 被引量:9
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作者 Qing-Sheng Yu Wan-Shou Guo +3 位作者 Li-Ming Cheng Yu-Feng Lu Jian-Ying Shen Ping Li 《Chinese Medical Journal》 SCIE CAS CSCD 2015年第14期1956-1963,共8页
Background: Appropriate expression and regulation of the transcriptome, which mainly comprise ofmRNAs and lncRNAs, are important for all biological and cellular processes including the physiological activities of bon... Background: Appropriate expression and regulation of the transcriptome, which mainly comprise ofmRNAs and lncRNAs, are important for all biological and cellular processes including the physiological activities of bone microvascular endothelial cells (BMECs). Through an intricate intraeellular signaling systems, the transcriptome regulates the pharmacological response of the cells. Although studies have elucidated the impact of glucocorticoids (GCs) cell-specific gene expression signatures, it remains necessary to comprehensively characterize the impact of lncRNAs to transcriptional changes. Methods: BMECs were divided into two groups. One was treated with GCs and the other left untreated as a paired control. Differential expression was analyzed with GeneSpring software V12.0 (Agilent, Santa Clara, CA, USA) and hierarchical clustering was conducted using Cluster 3,0 software. The Gene Ontology (GO) analysis was performed with Molecular Annotation System provided by CapitalBio Corporation. Results: Our results highlight the involvement of genes implicated in development, differentiation and apoptosis following GC stimulation. Elucidation of differential gene expression emphasizes the importance of regulatory gene networks induced by GCs. We identified 73 up-regulated and 166 down-regulated long noncoding RNAs, the expression of 107 of which significantly correlated with 172 mRNAs induced by hydrocortisone. Conclusions: Transcriptome analysis of BMECs from human samples was performed to identify specific gene networks induced by GCs. Our results identified complex RNA crosstalk underlying the pathogenesis of steroid-induced necrosis of femoral head. 展开更多
关键词 co-expression Network Intracellular Signaling Pathway Microvascular Endothelial Ceils Noncoding RNAs OSTEONECROSIS
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Screening biomarkers for spinal cord injury using weighted gene co-expression network analysis and machine learning 被引量:4
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作者 Xiaolu Li Ye Yang +3 位作者 Senming Xu Yuchang Gui Jianmin Chen Jianwen Xu 《Neural Regeneration Research》 SCIE CAS CSCD 2024年第12期2723-2734,共12页
Immune changes and inflammatory responses have been identified as central events in the pathological process of spinal co rd injury.They can greatly affect nerve regeneration and functional recovery.However,there is s... Immune changes and inflammatory responses have been identified as central events in the pathological process of spinal co rd injury.They can greatly affect nerve regeneration and functional recovery.However,there is still limited understanding of the peripheral immune inflammato ry response in spinal cord inju ry.In this study.we obtained microRNA expression profiles from the peripheral blood of patients with spinal co rd injury using high-throughput sequencing.We also obtained the mRNA expression profile of spinal cord injury patients from the Gene Expression Omnibus(GEO)database(GSE151371).We identified 54 differentially expressed microRNAs and 1656 diffe rentially expressed genes using bioinformatics approaches.Functional enrichment analysis revealed that various common immune and inflammation-related signaling pathways,such as neutrophil extracellular trap formation pathway,T cell receptor signaling pathway,and nuclear factor-κB signal pathway,we re abnormally activated or inhibited in spinal cord inju ry patient samples.We applied an integrated strategy that combines weighted gene co-expression network analysis,LASSO logistic regression,and SVM-RFE algorithm and identified three biomarke rs associated with spinal cord injury:ANO10,BST1,and ZFP36L2.We verified the expression levels and diagnostic perfo rmance of these three genes in the original training dataset and clinical samples through the receiver operating characteristic curve.Quantitative polymerase chain reaction results showed that ANO20 and BST1 mRNA levels were increased and ZFP36L2 mRNA was decreased in the peripheral blood of spinal cord injury patients.We also constructed a small RNA-mRNA interaction network using Cytoscape.Additionally,we evaluated the proportion of 22 types of immune cells in the peripheral blood of spinal co rd injury patients using the CIBERSORT tool.The proportions of naive B cells,plasma cells,monocytes,and neutrophils were increased while the proportions of memory B cells,CD8^(+)T cells,resting natural killer cells,rest 展开更多
关键词 bioinformatics analysis BIOMARKER CIBERSORT GEO dataset LASSO miRNA-mRNA network RNA sequencing spinal cord injury SVM-RFE weighted gene co-expression network analysis
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共表达SHMT和TPase载体的构建及双酶法合成L-色氨酸 被引量:9
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作者 李鑫 刘军 +1 位作者 赵沁沁 徐爱才 《生物工程学报》 CAS CSCD 北大核心 2010年第9期1302-1308,共7页
利用重组大肠杆菌表达丝氨酸羟甲基转移酶(SHMT)和色氨酸酶(TPase),并利用双酶法合成L-色氨酸。采用PCR从大肠杆菌K12基因组中扩增上述两种酶的基因,利用pET-28a载体,构建单表达重组质粒pET-SHMT、pET-TPase和共表达重组质粒pET-ST。将... 利用重组大肠杆菌表达丝氨酸羟甲基转移酶(SHMT)和色氨酸酶(TPase),并利用双酶法合成L-色氨酸。采用PCR从大肠杆菌K12基因组中扩增上述两种酶的基因,利用pET-28a载体,构建单表达重组质粒pET-SHMT、pET-TPase和共表达重组质粒pET-ST。将上述3种重组质粒转入大肠杆菌BL21(DE3)进行表达。SDS-PAGE结果表明,单表达基因工程菌BL21(DE3)/pET-SHMT和BL21(DE3)/pET-TPase分别在47kDa(SHMT)和50kDa(TPase)处有蛋白表达带;共表达基因工程菌BL21(DE3)/pET-ST在上述两处均有蛋白表达带。与宿主菌相比,单表达SHMT基因工程菌产酶活性提高了6.4倍;单表达TPase基因工程菌产酶活性提高了8.4倍;共表达SHMT和TPase基因工程菌产酶活性分别提高了6.1和6.9倍。利用工程菌所产酶进行双菌双酶法和单菌双酶法合成L-色氨酸。两菌双酶合成L-色氨酸的累积量达到41.5g/L,甘氨酸转化率为83.3%,吲哚转化率为92.5%;单菌双酶合成L-色氨酸的累积量达到28.9g/L,甘氨酸转化率为82.7%,吲哚转化率为82.9%。 展开更多
关键词 丝氨酸羟甲基转移酶 色氨酸酶 串联表达 双酶法合成
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卵巢上皮性肿瘤中PTEN和survivin的表达及意义 被引量:8
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作者 朱巧英 沈宇飞 +1 位作者 胡建铭 詹升华 《实用医学杂志》 CAS 北大核心 2009年第18期3009-3010,共2页
目的:研究卵巢上皮性肿瘤中PTEN和survivin的表达。方法:采用免疫组织化学法检测PTEN和survivin在57例卵巢上皮性癌、20例卵巢良性上皮性肿瘤和10例正常卵巢组织标本中的表达。结果:卵巢上皮性癌中survivin的阳性表达率(64.91%)明显高... 目的:研究卵巢上皮性肿瘤中PTEN和survivin的表达。方法:采用免疫组织化学法检测PTEN和survivin在57例卵巢上皮性癌、20例卵巢良性上皮性肿瘤和10例正常卵巢组织标本中的表达。结果:卵巢上皮性癌中survivin的阳性表达率(64.91%)明显高于正常卵巢组织(0%)及卵巢良性上皮性瘤(0%),P<0.01。卵巢上皮性癌中PTEN的阳性表达率(38.60%)明显低于正常卵巢组织(100%)及卵巢良性上皮性瘤(95.00%),P<0.05;survivin的阳性表达、PTEN的阴性表达及survivin阳性与PTEN阴性的共表达与卵巢上皮性癌的临床分期、病理分级正相关(P<0.05)。结论:Survivin的表达上调及PTEN的表达下调与卵巢上皮性癌的发生相关,并且在卵巢上皮性癌的发生发展中有协同作用,联合检测其中的指标对于评价卵巢上皮性癌的恶性程度、转移及预后具有临床价值。 展开更多
关键词 卵巢肿瘤 PTEN SURVIVIN 共表达
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Bcl-2、C-myc蛋白的表达与弥漫性大B细胞淋巴瘤预后的研究 被引量:8
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作者 廖银花 李爱平 +3 位作者 石旦 刘伶俏 陈亚辉 李晓杰 《诊断病理学杂志》 2020年第8期567-572,共6页
目的探讨Bcl-2、C-myc蛋白表达在弥漫性大B细胞淋巴瘤(DLBCL)诊断、预后判断及治疗中的价值。方法采用免疫组织化学染色法检测72例DLBCL组织标本中C-myc、Bcl-2、CD10、Bcl-6、MUM-1蛋白的表达情况。结果卡方检验统计分析:bcl-2在患者... 目的探讨Bcl-2、C-myc蛋白表达在弥漫性大B细胞淋巴瘤(DLBCL)诊断、预后判断及治疗中的价值。方法采用免疫组织化学染色法检测72例DLBCL组织标本中C-myc、Bcl-2、CD10、Bcl-6、MUM-1蛋白的表达情况。结果卡方检验统计分析:bcl-2在患者的临床分期、IPI评分及病理分型之间表达有显著差异(P<0.05);C-myc在患者的临床分期及IPI评分之间的表达有显著差异(P<0.05);bcl-2和C-myc蛋白共表达和非共表达在患者的临床分期、B症状及IPI评分之间有显著差异(P<0.05)。②Kaplan-Meier法单因素分析:患者的年龄、临床分期、B症状、IPI评分、病理分型、bcl-2蛋白表达及bcl-2/C-myc共表达与患者的平均OS及平均PFS比较有统计学意义(P<0.05)。患者的性别、原发部位、C-myc蛋白表达的平均OS和平均PFS比较无统计学意义(P> 0.05)。③COX多因素分析:DLBCL患者的年龄、临床分期、B症状和bcl-2蛋白与OS之间的分析有统计学意义(P<0.05);DLBCL患者的年龄、临床分期、B症状与PFS之间的分析有统计学意义(P<0.05)。结论 DLBCL患者的年龄、临床分期、有无B症状、IPI评分、病理分型和bcl-2蛋白可以初步判断预后,对指导临床治疗有一定意义;患者的年龄、临床分期、B症状和Bcl-2蛋白的表达是影响DLBCL患者预后的独立危险因素;C-myc的单独表达尚不能作为影响DLBCL患者预后的独立危险因素。 展开更多
关键词 弥漫大B细胞性淋巴瘤 BCL-2 C-MYC 共表达 预后
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肝癌靶向性SEA-CD80基因共表达重组腺病毒载体的构建及表达鉴定 被引量:8
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作者 司少艳 隋延仿 +4 位作者 胡沛臻 张秀敏 葛伟 李增山 黄杨 《细胞与分子免疫学杂志》 CAS CSCD 北大核心 2006年第5期613-616,共4页
目的:构建肝癌靶向性葡萄球菌肠毒素A(SEA)和CD80基因共表达重组腺病毒载体。方法:首先利用现有的腺病毒穿梭质粒pShuttle和pShuttleCMV,构建新的不带CMV增强子/启动子而带有polyA加尾信号穿梭质粒,命名为pShuttle2。将AFP增强子、启动... 目的:构建肝癌靶向性葡萄球菌肠毒素A(SEA)和CD80基因共表达重组腺病毒载体。方法:首先利用现有的腺病毒穿梭质粒pShuttle和pShuttleCMV,构建新的不带CMV增强子/启动子而带有polyA加尾信号穿梭质粒,命名为pShuttle2。将AFP增强子、启动子、SEA及CD80基因分别从已构建的pKSEP载体和pMD18TBIS载体上,分别亚克隆至pShuttle2中,再与腺病毒骨架质粒pAdEasy1共转化E.coliBJ5183。以获得的重组子转染HEK293细胞后制备重组腺病毒,然后感染高表达AFP的肝癌细胞系Hepa16和不表达AFP的黑色素瘤细胞系B16、成纤维细胞系NIH3T3。采用间接免疫荧光法,激光共聚焦显微镜观察和流式细胞术检测SEA和CD80在细胞膜表面的表达。采用3H掺入法检测膜表达的SEA诱导淋巴细胞增殖的活性。结果:以制备的重组腺病毒感染肿瘤细胞后,SEA和CD80能够靶向性地共表达在高表达AFP的Hepa16细胞膜上,而在不表达AFP的B16、NIH3T3细胞膜上不表达。结论:成功地构建肝癌靶向性SEA和CD80基因共表达重组腺病毒载体,为进一步研究SEA和CD80在肝癌靶向基因治疗中的联合应用及其抗肿瘤免疫机制奠定了基础。 展开更多
关键词 葡萄球菌肠毒素A 超抗原 CDSO 共表达 腺病毒
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共表达猪呼吸与繁殖障碍综合征病毒NJ-a株ORF4、ORF5与ORF6基因重组改良型痘苗病毒安卡拉株的构建 被引量:8
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作者 郑其升 毕志香 +2 位作者 李鹏 陈德胜 陈溥言 《微生物学报》 CAS CSCD 北大核心 2007年第2期345-349,共5页
为探讨共表达猪繁殖与呼吸综合征病毒(Porcinerep roductive and respiratory syndrome Virus,PRRSV)保护性抗原基因的重组改良型痘苗病毒安卡拉株(Modified Vaccinia Virus Ankala,MVA)的免疫效力,将PRRSVNJ-a株ORF4、ORF5和ORF6基因... 为探讨共表达猪繁殖与呼吸综合征病毒(Porcinerep roductive and respiratory syndrome Virus,PRRSV)保护性抗原基因的重组改良型痘苗病毒安卡拉株(Modified Vaccinia Virus Ankala,MVA)的免疫效力,将PRRSVNJ-a株ORF4、ORF5和ORF6基因插入转移载体pⅡLR中,获得了三基因共表达的转移载体pⅡLR-ORF5/ORF6/ORF4,通过同源重组的方法获得重组病毒rMVA-GP5/M/GP4。以lacZ为报告基因进行噬斑筛选和重组病毒纯化后,PCR方法证明ORF4、ORF5和ORF6成功的插入MVA基因组中;经Western blot检测与间接免疫荧光试验证实,重组病毒感染细胞能正确表达PRRSVGP4、GP5与M蛋白。用rMVA-GP5/M/GP4免疫6周龄Babl/C小鼠,首免后3周可检测到特异性PRRSV中和抗体,8周后中和抗体效价可达25,并能继续维持4周;淋巴细胞增殖试验结果表明,重组病毒免疫小鼠产生强烈的特异性细胞增殖反应。上述研究结果表明rMVA-GP5/M/GP4具有良好的免疫原性,可作为预防PRRS的候选疫苗进一步研究。 展开更多
关键词 猪繁殖与呼吸综合征病毒 修饰的痘苗病毒安卡拉株 GP4 GP5与M蛋白 共表达
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分子伴侣共表达对低温脂肪酶Lip-837异源可溶性表达的影响 被引量:7
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作者 崔硕硕 张镱 +1 位作者 林学政 沈继红 《海洋科学进展》 CAS CSCD 北大核心 2011年第1期105-112,共8页
将南极嗜冷菌Moritellasp.2-5-10-1的低温脂肪酶基因Lip-837克隆到表达载体pColdⅢ中,成功构建了重组质粒pColdⅢ+Lip-837,并转化大肠杆菌E.coli BL21(DE3)。SDS-PAGE实验结果表明,实现了该基因在E.coli的异源表达,且其表达量达到细胞... 将南极嗜冷菌Moritellasp.2-5-10-1的低温脂肪酶基因Lip-837克隆到表达载体pColdⅢ中,成功构建了重组质粒pColdⅢ+Lip-837,并转化大肠杆菌E.coli BL21(DE3)。SDS-PAGE实验结果表明,实现了该基因在E.coli的异源表达,且其表达量达到细胞总蛋白的39%;但表达产物主要以包涵体形式存在。将该重组质粒pColdⅢ+Lip-837分别与不同分子伴侣质粒,如pGro7,pKJE7,pG-Tf2,pTf16在E.coli BL21(DE3)中实现了共表达。SDS-PAGE结果表明,4种分子伴侣均能明显提高目的蛋白—脂肪酶Lip-837的可溶性表达,但不同分子伴侣提高可溶性蛋白表达的程度有所不同,如仅表达pColdⅢ+Lip-837时,低温脂肪酶可溶性蛋白占总表达量的比例为5%;当与分子伴侣pGro7共表达时,对脂肪酶可溶性蛋白表达提高的比例最大,其可溶性酶蛋白的比例可占总表达量的42%;分子伴侣质粒pG-Tf2促进可溶性表达的效果较差,其可溶性酶蛋白的比例仅占其总表达量的16.6%。 展开更多
关键词 低温脂肪酶 分子伴侣 共表达 可溶性表达
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光皮桦MYB基因的克隆及表达和调控分析 被引量:7
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作者 倪飞 励文豪 +2 位作者 林二培 童再康 黄华宏 《林业科学》 EI CAS CSCD 北大核心 2018年第12期70-81,共12页
【目的】MYB转录因子是植物中一类重要的转录因子,在调控次生壁纤维素和木质素等的合成中具有重要作用。为了挖掘参与光皮桦木材形成的MYB基因,本研究通过克隆MYB基因,分析其序列特征、表达模式和下游调控基因,以期为后续功能深入解析... 【目的】MYB转录因子是植物中一类重要的转录因子,在调控次生壁纤维素和木质素等的合成中具有重要作用。为了挖掘参与光皮桦木材形成的MYB基因,本研究通过克隆MYB基因,分析其序列特征、表达模式和下游调控基因,以期为后续功能深入解析和分子辅助育种提供理论依据和基因资源。【方法】利用RACE技术分离到4个光皮桦BlMYB基因的cDNA片段。利用生物信息学工具对这些基因的序列特征进行分析,利用实时荧光定量PCR分析BlMYB及预测的下游基因在不同器官(雌花序、雄花序、嫩芽、嫩叶、成熟叶、茎)、组织(内外层木质部、韧皮部、形成层)中,以及应拉木诱导早期阶段的表达差异。以2个木质部特异表达的BlMYB为指导基因,采用相互排名法和Cytoscape软件构建共表达网;使用Plant CARE在线查询共表达的下游基因启动子区元件。【结果】分离到4个BlMYB的全长cDNA序列,分别命名为BlMYB1、BlMYB2、BlMYB3和BlMYB4,它们编码的蛋白分别由395、252、258、320个氨基酸残基组成,且在靠近N端都有R2R3结构域。4个BlMYB氨基酸序列间一致性27%~37%,而与它们同源的拟南芥AtMYB氨基酸序列间的一致性为39%~55%。4个BlMYB基因都含有1~2个内含子。进化树构建分析发现4个BlMYB分属于4个不同的分支,其中BlMYB2、BlMYB4分别与细胞次生壁形成相关的MYB聚在一起。BlMYB1、BlMYB3在成熟叶片中优势表达,且随叶片成熟表达水平呈递增趋势,可能与叶片的发育有关。BlMYB2在木质化茎段的木质部强烈表达,而在叶片、韧皮部等组织中的表达相对较弱;在应拉木诱导形成的早期阶段,应拉木中BlMYB2呈下调表达,尤其是拉弯处理48 h和7天时的相应数值均显著低于直立木。BlMYB4在茎的木质部、成熟雄花序中优势表达,在根、嫩叶中的表达水平较低;同时,在应拉木诱导形成早期阶段,BlMYB4在应拉木和对应木中分别呈现上� 展开更多
关键词 光皮桦 BlMYB基因 应拉木 表达模式 共表达
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Mining the key regulatory genes of chicken inosine 5'-monophosphate metabolism based on time series microarray data 被引量:6
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作者 Teng Ma Lu Xu +4 位作者 Hongzhi Wang Jing Chen Lu Liu Guobin Chang Guohong Chen 《Journal of Animal Science and Biotechnology》 SCIE CAS CSCD 2015年第3期280-290,共11页
IMP (inosine 5'-monophosphate) is a compound that enhances the flavor of poultry meat. IMP has become a new breeding trait to improve poultry meat quality. We tried to identify several potential regulatory genes, a... IMP (inosine 5'-monophosphate) is a compound that enhances the flavor of poultry meat. IMP has become a new breeding trait to improve poultry meat quality. We tried to identify several potential regulatory genes, and construct their predicted regulatory relationships. Time series gene expression profiles of thigh muscle tissues of Rugao chicken, a famous indigenous breed in China, were performed for analysis of genes that are co-expressed or correlated with the concentration of IMP. We found 15 crucial co-expression genes, which are Hspa2, Pten, Gabpa, Bpi, Mkll, Srf,, Cd34, Hspa4, EtvS, Bmpr2, Gdel, IgfbpS, Cd28, Pecam1 and Gja1, that may directly or indirectly regulate IMP metabolism. Eventually, we computed the correlation coefficient between 19 IMP Genes and 15 CGs (15 co-expression genes), and we identified and constructed a predicted regulation network. In conclusion, variation of IMP concentration was primarily connected with the muscle development process. During this process, 15 CGs were identified that may have significant influence on IMP metabolism. In particular, Bmpr2, Pten and co-expression genes correlated with Entpd8 might play important roles in regulating IMP de novo synthesis, decomposition and salvage synthesis. 展开更多
关键词 co-expression Hub genes IMP metabolism Regulation network
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