检索结果-南通市图书馆

维普期刊数据库

梅花鹿致敏与休眠鹿茸干细胞差异表达蛋白的2d-dige分析

在线全文

学校读者我要写书评

暂无评论

梅花鹿致敏与休眠鹿茸干细胞差异表达蛋白的2D-DIGE分析

作者：董振中国农业科学院

学位级别：硕士

鹿茸是目前已知的唯一一个可周期性再生的哺乳动物附属器官,实验证明是角柄骨膜中的角柄骨膜细胞具有完全再生出鹿茸组织的能力。根据与皮肤接触紧密程度的不同,角柄骨膜可以分为致敏区与休眠区,前者与其皮肤的相互作用能使角柄致敏,最... 详细信息

鹿茸是目前已知的唯一一个可周期性再生的哺乳动物附属器官,实验证明是角柄骨膜中的角柄骨膜细胞具有完全再生出鹿茸组织的能力。根据与皮肤接触紧密程度的不同,角柄骨膜可以分为致敏区与休眠区,前者与其皮肤的相互作用能使角柄致敏,最终使角柄骨膜细胞,即鹿茸干细胞,发育成鹿茸组织。因此,角柄骨膜干细胞中的蛋白质组的变化可能在触发鹿茸再生的过程中扮演了主要角色。以往,对角柄骨膜致敏区与休眠区(尚未与覆盖皮肤建立相互作用)的研究以定性蛋白质组学技术为主,如双向电泳技术;而通过定量蛋白质组学技术对此方面进行的研究尚未见诸报道。本研究首先将梅花鹿(Cervus nippon)角柄骨膜细胞作为试验材料,从细胞破碎方法、染色方法、等电聚焦(Isoelectric Focusing,IEF)总伏小时数(volt-hours)和蛋白提取纯化组合4个不同的角度来优化双向电泳过程,为后续基于双向电泳原理的荧光差异显示双向电泳技术(Two-dimensional Fluorescence difference in Gel Electrophoresis,2d-dige)所进行的致敏与休眠鹿茸干细胞差异蛋白质组学研究提供借鉴依据。结果显示:与超声相比,Bullet Blender对于细胞样品的破碎效果更好;双染法能够得到更多且更清晰的蛋白点;IEF总伏小时数为15000时所得到的凝胶图谱最适合于后续软件分析;通过对6种蛋白提取与纯化组合的比较来看,自制裂解液与2d Clean-up Kit处理可以得到更清晰且具有更多蛋白丰度信息的凝胶图谱。随后采用荧光差异显示双向电泳技术分离蛋白样品;利用deCyder 2d 7.2分析软件对2d-dige所得到的凝胶图谱进行统计分析以寻找致敏与休眠鹿茸干细胞的差异表达蛋白;MALdI-TOF-MS(Matrix-assisted Laser desorption/Ionization Time-of-Flight Mass Spectrometry)鉴定差异蛋白,通过Mascot软件搜索NCBInr数据库寻找匹配蛋白;采用PANTHER软件对差异蛋白进行聚类分析,REACTOME数据库分析差异蛋白所参与的信号通路。结果显示:在致敏鹿茸干细胞与休眠鹿茸干细胞蛋白丰度的比较中,共159个蛋白点比值≥1.1倍或≤-1.1倍(p27种蛋白质。并对已鉴定到的蛋白进行了GO分析和信号通路富集等一系列生物信息学分析。通过对质谱数据的归纳分析后发现:POTE ankyrin domain family member E、SUMO-activating enzyme subunit 1、Aldolase B、Glucose-regulated peptide 78、Inositol 1,4,5-trisphosphate receptor type1、Tetratricopeptide repeat protein 36、Histone H4、ATP synthase subunit beta和Vimentin等九种蛋白可能与触发鹿茸再生有密切的关系。其中,后四种蛋白是deCyder 2d 7.2分析软件所判定的Marker蛋白,这些蛋白可能是决定致敏与休眠鹿茸干细胞再生功能有无的重要调控蛋白。综上可知,鹿茸再生是鹿茸干细胞从休眠到致敏的转化过程,需要多种蛋白分子以及信号通路的综合调控。本研究为深入探讨鹿茸所具有的独特再生分子调控机制提供了理论依据。

关键词：角柄骨膜鹿茸干细胞再生双向电泳 2d-dige

同方学位论文库评论

在线全文

同方学位论文库

学校读者我要写书评

暂无评论

2d-dige技术在蛋白质组学中的应用

生物学杂志 2011年第3期28卷 84-87,96页

作者：杜俊变王丽惠段江燕山西师范大学生命科学学院山西临汾041000 海南大学材料与化工学院生物工程系海口570228

双向荧光差异凝胶电泳(2d-d IGE)作为一种新型的蛋白质组分析技术,已经被广泛应用于动物、植物、微生物以及人类差异蛋白的研究。在动物医学方面,采用dige技术,通过对不同类型,不同个体的细胞、组织、或经过不同处理和不同生长条件下蛋... 详细信息

双向荧光差异凝胶电泳(2d-d IGE)作为一种新型的蛋白质组分析技术,已经被广泛应用于动物、植物、微生物以及人类差异蛋白的研究。在动物医学方面,采用dige技术,通过对不同类型,不同个体的细胞、组织、或经过不同处理和不同生长条件下蛋白质表达差异分析,在研究疾病的分子机理、分子诊断、药物作用机理、毒理学等方面都有广泛的应用。在植物学方面,该技术可以用来分离和分析植物亚细胞结构蛋白质组以及在生物和非生物胁迫下,植物细胞中蛋白质表达的差异性,从而建立差异蛋白相互作用网络图,从而为研究伤害机制提供一定的依据。

关键词： 2d-dige 蛋白质组

维普期刊数据库

梅花鹿致敏与休眠鹿茸干细胞差异蛋白表达的2d-dige分析

在线全文

学校读者我要写书评

暂无评论

梅花鹿致敏与休眠鹿茸干细胞差异蛋白表达的2D-DIGE分析

Proteomic Study of differentially Expressed Proteins in Seeds between Parents and Offspring of Castor Bean(Ricinus communis L.)

作者：董振王权威刘振孙红梅李春义中国农业科学院特产研究所特种动物分子生物学国家重点实验室

【目的】对梅花鹿(Cervus nippon)致敏鹿茸干细胞与休眠鹿茸干细胞表达蛋白进行差异筛选、鉴定及生物信息分析,为深入探讨鹿茸独特的再生分子调节机制奠定基础。【方法】采用双向荧光差异凝胶电泳(Two-dimensional fluorescence differe... 详细信息

【目的】对梅花鹿(Cervus nippon)致敏鹿茸干细胞与休眠鹿茸干细胞表达蛋白进行差异筛选、鉴定及生物信息分析,为深入探讨鹿茸独特的再生分子调节机制奠定基础。【方法】采用双向荧光差异凝胶电泳(Two-dimensional fluorescence difference in gel electrophoresis,2d-dige)分离蛋白样品;利用deCyder 7.2分析软件对2d-dige图像进行统计学分析寻找差异表达蛋白;MALdI-TOF-MS(Matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry)鉴定差异蛋白,通过Mascot软件搜索NCBInr数据库寻找匹配的蛋白;采用PANTHER(Protein Analysis Through Evolutionary Relationships)软件对差异蛋白进行聚类分析,REACTOME数据库分析差异蛋白所参与的信号通路。【结果】得到了致敏鹿茸干细胞与休眠鹿茸干细胞2d-dige图谱,致敏鹿茸干细胞与休眠鹿茸干细胞蛋白丰度相比较比值≥1.1倍以及比值≤-1.1倍(PdA(Extended data analysis)分析得到了多个Marker蛋白,质谱鉴定了84个差异蛋白质点,48个为阳性结果,共来自27种蛋白质。并对已鉴定蛋白进行了GO分析以及信号通路富集分析。【结论】致敏鹿茸干细胞与休眠鹿茸干细胞蛋白差异明显,质谱鉴定获得了来自多种可能与鹿茸再生相关的差异蛋白。由此可知,鹿茸再生是鹿茸干细胞从休眠到致敏的转化过程,需要多种蛋白分子以及信号通路的综合调控。

关键词：鹿茸干细胞再生蛋白质组学 2d-dige

来源： cnki会议评论

在线全文

cnki会议

学校读者我要写书评

暂无评论

Phyton-International Journal of Experimental Botany 2023年第6期92卷 1765-1792页

作者： Xiaotian Liang Qi Wen Rui Luo Yanxin Zhang Mingda Yin Yanpeng Wen Xuemei Hu Zhiyan Wang Yumiao Huo Fenglan Huang College of Life Science and Food Engineering Inner Mongolia Minzu UniversityTongliao028000China Key Laboratory of Castor Breeding of State Ethnic Affairs Commission Tongliao028000China Research Center for Engineering and Technology of Castor Industry by Universities in Inner Mongolia Autonomous Region Tongliao028000China Key Laboratory of Castor Breeding of Inner Mongolia Autonomous Region Tongliao028000China Collaborative Innovation Center of Castor Industry of Inner Mongolia Autonomous Region Tongliao028000China Engineering Research Center of Castor Industry Technology Innovation Inner Mongolia Autonomous RegionTongliao028000China

Castor bean(Ricinus communis L.),is one of the top 10 oilseed crops in the world and,therefore,of high economic *** is one of the most effective ways to breed new varieties with high yield,high oil content,and better stress ***,prediction of desired traits in castor hybrid offspring is particularly *** this study,proteomic analysis was performed to identify differentially expressed proteins(dEPs)in seeds between castor hybrid offspring and their female(Lm female line aLmAB2)and male parents(CSR·181).Among the dEPs upregulated in the seeds of hybrid offspring,the majority were related to seed yield and stress tolerance,while some were related to oil synthesis and fatty acid synthesis and metabolism in *** other words,the hybrid offspring showed heterosis for seed yield,stress tolerance,oil synthesis,and fatty acid synthesis and metabolism when compared with their ***,real-time quantitative polymerase chain reaction assays were performed on 12 genes encoding dEPs involved in oil synthesis,pollen abortion,yield,and stress tolerance of *** results showed that the expression levels of the 12 genes were consistent with those of the dEPs.

关键词： Castor bean 2d-dige heterosis

Effect of high-nitrogen fertilizer on gliadin and glutenin subproteomes during kernel development in wheat(Triticum aestivum L.)

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

The Crop Journal 2020年第1期8卷 38-52页

作者： Shoumin Zhen Xiong deng Xuexin Xu Nannan Liu dong Zhu Zhimin Wang Yueming Yan College of Life Science Capital Normal UniversityBeijingChina Hubei Collaborative Innovation Center for Grain Industry(HCICGI) Yangtze University434025 JingzhouChina Institute of Scientific and Technical Information of China Beijing 100038China College of Agronomy and Biotechnology China Agricultural University100081 BeijingChina

Nitrogen(N),a macronutrient essential for plant growth and development,is needed for biosynthesis of protein and starch,which affect grain yield and *** of high-N fertilizer increases plant growth,grain yield,and flour *** this study,we performed the first comparative analysis of gliadin and glutenin subproteomes during kernel development in the elite Chinese wheat cultivar Zhongmai 175 under high-N conditions by reversed-phase ultra-performance liquid chromatography and twodimensional difference gel electrophoresis(2d-dige).Application of high-N fertilizer led to significant increases in gluten macropolymer content,total gliadin and glutenin content,and the accumulation of individual storage protein *** 126 differentially accumulated proteins(dAPs)induced by high-N conditions,24 gliadins,12 high-molecularweight glutenins,and 27 low-molecular-weight glutenins were significantly *** during five kernel developmental stages displayed multiple patterns of *** particular,gliadins and glutenins showed respectively five and six accumulation *** accumulation of storage proteins under high-N conditions may lead to improved dough properties and bread quality.

关键词： Bread wheat High nitrogen Gliadins Glutenins RP-UPLC 2d-dige

维普期刊数据库

Identification of transporter proteins for PQQ-secretion pathways by transcriptomics and proteomics analysis in Gluconobacter oxydans WSH-003

在线全文

学校读者我要写书评

暂无评论

Frontiers of Chemical Science and Engineering 2017年第1期11卷 72-88页

作者： Hui Wan Yu Xia Jianghua Li Zhen Kang Jingwen Zhou School of Biotechnology and Key Laboratory of Industrial B iotechnology Ministry of Education Jiangnan University Wuxi 214122 China Synergetic Innovation of Center of Food Safety and Nutrition Jiangnan University Wuxi 214122 China

Pyrroloquinoline quinone （PQQ） plays a sig- nificant role as a redox cofactor in combination with dehydrogenases in bacteria. These dehydrogenases play key roles in the oxidation of important substrates for the biotechnology industry, such as vitamin C production. While biosynthesis of PQQ genes has been widely studied, PQQ-transport mechanisms used both two-dimensional remain unclear. Herein, we fluorescence-difference gel electrophoresis tandem mass spectrometry and RNA sequencing to investigate the effects ofpqqB overexpres- sion in an industrial strain of Gluconobacter oxydans WSH-003. We have identified 73 differentially expressed proteins and 99 differentially expressed genes, a majority of which are related to oxidation-reduction and transport processes by gene ontology analysis. We also described several putative candidate effectors that responded to increased PQQ levels resulting from pqqB overexpression. Furthermore, quantitative PCR was used to verify five putative PQQ-transport genes among different PQQ producing strains, and the results showed that ompW, B932 1930 and B932_2186 were upregulated in all conditions. Then the three genes were over-expressed in G. oxydans WSH-003 and PQQ production were detected. The results showed that extracellular PQQ of B932_1930 （a transporter） and B932_2186 （an ABC transporter perrnease） overexpression strains were enhanced by 1.77- fold and 1.67-fold, respectively. The results suggest that the proteins encoded by PqqB, B932_1930 and B932 2186 might enhance the PQQ secretion process.

关键词： 2d-dige pqqB pyrroloquinoline quinone,RNA-Seq Vitamin C

Comparative proteomics analysis of lungs artificially infected with H-PRRSV and N-PRRSV by Two-dimensional fluorescence difference gel electrophoresis

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Comparative proteomics analysis of lungs artificially infect...

Comparative Proteomic Analysis of differentially Expressed Proteins Beween U251 Cells Transfected by pcdNA3.1（+） or LRRC4 gene

第十五次全国动物遗传育种学术讨论会

作者： Jianyu Jia,Shuqi Xiao,Qiwei Wang,Yaosheng Chen~* State Key Laboratory of Biocontrol,School of Life Sciences,Sun Yat-sen University,Guangzhou 510006

Introduction Porcine reproductive and respiratory syndrome with PRRS virus(PRRSV) infection,being one of the most economically important diseases affecting swine industry worldwide,causes significant economic losses e... 详细信息

关键词： PRRS H-PRRSV N-PRRSV Lungs Pathogenesis 2d-dige

来源： cnki会议评论

在线全文

cnki会议

学校读者我要写书评

暂无评论

Comparative Proteomic Analysis of Differentially Expressed P...

水稻草状矮化病毒侵染寄主水稻差异表达蛋白的鉴定和分析

2008年中国病理生理学会第十一届肿瘤和第十二届免疫专...

作者： Minghua Wu Jing Yang Zuping Zhang Guiyuan Li Cancer Research Institute,Central South University,110# Xiang-Ya Road,Changsha,410078,Hunan,The People''s Republic of China

＜正＞Leucine-rich repeats containing 4 （LRRC4） is a specially expressed gene in brain,and also a tumor suppressive gene of ***4 is a functional gene in neural development and axon growth,and associated with glioma grade ***4 expression is gradually reduced, even absent accompany with glioma grade *** reexpression of LRRC4 can inhibit U251 cells proliferation,adhesion,chemotatic effect and *** further understand the regulating effect of LRRC4 on proteins in glioma,differential in-gel electrophoresis（dige） and mass spectrometry（MS） were used to gain the differentially expressed proteins between U251 cells transfected by pcdNA3.1（+） or LRRC4 *** these differentially expressed proteins were mainly involved in diverse biology events,such as cell Metabolism,proliferation,translation,signal transduction et *** data indicated that LRRC4 may regulate the above differentially expressed proteins to participate in initiation and progression of glioma.

关键词： LRRC4 gliobalstoma 2d-dige MALdI-TOF/TOF-MS/MS

来源： cnki会议评论

在线全文

cnki会议

学校读者我要写书评

暂无评论

中国农业科学 2014年第9期47卷 1725-1734页

作者：丁新伦谢荔岩吴祖建福建农林大学植物病毒研究所福建省植物病毒重点实验室

【目的】对水稻草状矮化病毒(Rice grassy stunt virus,RGSV)侵染寄主水稻后其叶片的差异表达蛋白进行筛选、鉴定和生物信息学分析,为进一步研究RGSV和寄主水稻互作的分子机制提供线索。【方法】采用双向荧光差异凝胶电泳(Two-dimension... 详细信息

【目的】对水稻草状矮化病毒(Rice grassy stunt virus,RGSV)侵染寄主水稻后其叶片的差异表达蛋白进行筛选、鉴定和生物信息学分析,为进一步研究RGSV和寄主水稻互作的分子机制提供线索。【方法】采用双向荧光差异凝胶电泳(Two-dimensional fluorescence difference in gel electrophoresis,2d-dige)和Image Master 2d platinum 7.0分析软件寻找差异表达蛋白,MALdI-TOF-MS(Matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry)鉴定差异蛋白,利用BioTools软件搜索NCBI数据库,寻找匹配的相关蛋白质和功能查询。采用GOminer软件对差异蛋白进行GO(Gene ontology)聚类分析,KEGG(Kyoto encyclopedia of genes and genomes)数据库分析差异蛋白所参与的生物通路。【结果】建立了RGSV侵染与未侵染水稻的叶片双向荧光差异凝胶电泳图谱,RGSV病株与健株相比,差异倍数大于1.5的差异蛋白质点有173个,其中表达丰度升高的点有72个,表达丰度下降的点有101个,质谱鉴定了44个差异蛋白质点,25个获得成功鉴定,分属于24种蛋白质,包括RGSV的2种蛋白20.6K非结构蛋白和P5蛋白,水稻中的蛋白推定的核酮糖-1,5-二磷酸羧化酶/加氧酶大亚基、景天庚酮糖-1,7-二磷酸酶前体、推定的酪氨酸磷酸酶、HAd超家族水解酶-亚族IA蛋白变体3蛋白、水稻核酮糖-1,5-二磷酸羧化酶/加氧酶小亚基结合2-羧基阿拉伯糖醇-1,5-二磷酸复合体、类似C1结构域蛋白以及其他一些功能未知蛋白和假定蛋白。对已鉴定蛋白的生物信息分析显示,差异蛋白涉及氮素固定、氮循环代谢过程、含氮化合物代谢过程、单一生物体代谢过程、代谢过程和生物过程等6个生物学过程,在生物功能上分属7类,包括分子功能、酸胺连接酶活性、酰胺连接酶活性、催化活性、谷氨酸氨连接酶、连接酶活性和形成C-N键的连接酶活性,在细胞组件上,差异蛋白分布于不同的细胞位置,涉及细胞组分、细胞器、细胞、膜结合细胞器、囊(泡)、细胞部分、膜结合囊(泡)、胞内、胞内部分、胞内细胞器、细胞质、胞内膜结合细胞器、细胞质部分、质体、细胞质囊(泡)和细胞质膜结合囊(泡)。KEGG通路分析显示,差异蛋白参与了代谢途径、光合生物固碳反应、碳代谢、乙醛酸和二羧酸代谢、丙氨酸、天门冬氨酸和谷氨酸代谢、氨基酸生物合成、丙酮酸盐代谢、精氨酸和脯氨酸代谢、谷胱甘肽代谢和氮代谢途径等10个KEGG代谢通路。【结论】RGSV侵染诱导了水稻差异蛋白质表达,获得了一些与RGSV侵染水稻相关的蛋白质分子,差异蛋白涉及到多个功能类别。其中与氮有关的生物学过程和光合作用过程变化较为明显。

关键词：水稻草状矮化病毒差异表达蛋白 2d-dige MALdI-TOF-MS