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The Role of Linker Histone Mutation in Oncogenesis: Molecular Mechanism and Structural Impact
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作者 Gege Liu Houfang Zhang Yunhui Peng 《BIOCELL》 2025年第4期519-538,共20页
Nucleosomes play a vital role in chromatin organization and gene regulation,acting as key hubs that inter-act with various chromatin-associated factors through diverse binding mechanisms.Recent research has highlighte... Nucleosomes play a vital role in chromatin organization and gene regulation,acting as key hubs that inter-act with various chromatin-associated factors through diverse binding mechanisms.Recent research has highlighted the prevalence of mutations in linker histones across different types of cancer,emphasizing their critical involvement in cancer progression.These cancer-associated mutations in linker histones have been shown to disrupt nucleosome stacking and the formation of higher-order chromatin structures,which in turn significantly affect epigenetic regulatory processes.In this review,we provide a comprehensive analysis of how cancer-associated linker histone mutations alter their physicochemical properties,influencing their binding to nucleosomes,and overall chromatin architecture.Additionally,we explore the significant impact of mutations near post-translational modification sites,which further modulate chromatin dynamics and regulatory functions,offering insights into their role in oncogenesis and potential therapeutic targets. 展开更多
关键词 linker histone H1 EPIGENETICS histone cancer mutations chromatin structure NUCLEOSOME
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1.42-fold enhancement of formate selectivity by linker conversion on the Zn-based metal organic framework catalyst 被引量:1
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作者 Yayu Guan Yuyu Liu +2 位作者 Fanghua Ning Jin Yi Jiujun Zhang 《Journal of Energy Chemistry》 SCIE EI CAS CSCD 2024年第3期183-190,I0006,共9页
Electro-reduction of carbon dioxide(ERCO_(2)) is considered an effective method to alleviate the greenhouse effect and produce value-added chemicals.Achieving the dominant selectivity of Zn-based catalysts for formate... Electro-reduction of carbon dioxide(ERCO_(2)) is considered an effective method to alleviate the greenhouse effect and produce value-added chemicals.Achieving the dominant selectivity of Zn-based catalysts for formate remains a challenge.In this article,the ZnIn-E_(12) catalyst is successfully prepared by solvent assisted ligand exchange(SALE) method to convert organic ligands,achieving a Faradaic efficiency of 72.28% for formate at-1.26 V vs.RHE(V_(RHE)),which is 1.42 times higher than the original catalyst.Evidence shows that the successful conversion of organic ligands can transform the catalyst from the original large size polyhedron to cross-linked network of particles with a diameter of about 30 nm.The increased specific surface area can expose more active sites and facilitate the electrocatalytic conversion of CO_(2) to formate.This work is expected to provide inspiration for the regulation of formate selectivity and catalyst size in Zn-based catalysts. 展开更多
关键词 ELECTROCATALYST Carbon dioxide FORMATE linker conversion
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Alkyl-thiophene-alkyl linkers to construct double-cable conjugated polymers for single-component organic solar cells 被引量:1
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作者 Wenbin Lai Safakath Karuthedath +4 位作者 Chengyi Xiao Lei Meng Frédéric Laquai Weiwei Li Yongfang Li 《Chinese Chemical Letters》 SCIE CAS CSCD 2024年第1期542-546,共5页
In this work,semirigid linkers of the alkyl-thiophene-alkyl structure are developed to construct double-cable polymers.Three alkyl units,propyl(C3H6),hexyl(C6H12),and dodecyl(C12H24),are applied as semirigid linkers,y... In this work,semirigid linkers of the alkyl-thiophene-alkyl structure are developed to construct double-cable polymers.Three alkyl units,propyl(C3H6),hexyl(C6H12),and dodecyl(C12H24),are applied as semirigid linkers,yielding three double-cable polymers:PBC6-T,PBC12-T,and PBC24-T,respectively.PBC12-T which uses C6H12-thiophene-C6H12 linkers is found to exhibit the best device efficiency of 5.56%,while PBC6-T and PBC24-T with shorter or longer linkers yield device efficiencies of only 2.65%and 1.09%in single-component organic solar cells(SCOSCs).Further studies reveal that PBC12-T exhibits higher crystallinity and improved charge transport,resulting in better efficiencies.Our work provides an approach to construct double-cable conjugated polymers with long alkyl linkers,and it shows the importance of the linker length for the photovoltaic performance of SCOSCs. 展开更多
关键词 Double-cable conjugated polymer Single-component organic solar cell Crystallinity Semirigid linkers Alkyl-thiophene-alkyl
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Near-infrared double-cable conjugated polymers based on alkyl linkers with tunable length for single-component organic solar cells
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作者 Yikun Wang Qiaomei Chen +4 位作者 Shijie Liang Dongdong Xia Chaowei Zhao Christopher R.McNeill Weiwei Li 《Chinese Chemical Letters》 SCIE CAS CSCD 2024年第4期452-456,共5页
The photovoltaic properties of double-cable conjugated polymers are significantly influenced by the length of the alkyl linkers that connect donor backbones and acceptor side units. In this study, a series of 2-(3-oxo... The photovoltaic properties of double-cable conjugated polymers are significantly influenced by the length of the alkyl linkers that connect donor backbones and acceptor side units. In this study, a series of 2-(3-oxo-2,3-dihydroinden-1-ylidene)malononitrile(IC)-based double-cable polymers with alkyl linkers ranging from C_8H_(16)to C_(16)H_(32)(Px, x = 8, 10, 12, 14, 16) were synthesized for single-component organic solar cells(SCOSCs). Among these, the linker length x = 12(P12) is found to optimize the power conversion efficiencies(PCEs) in SCOSCs. Specifically, PCEs increase from P8 to P12 and then decline from P12to P16. Detailed investigations of optical absorption, charge transport, and morphology provide insights into the underlying factors contributing to these PCE variations. The findings indicate that the exceptional photovoltaic properties observed in P12 can be attributed to three key factors: A delicate balance between enhanced charge separation facilitated by the increased spacer length and reduced crystallinity resulting from longer spacers, higher charge mobilities, and well-balanced hole/electron transport characteristics. This study highlights the critical role of linker length in determining the photovoltaic properties of double-cable conjugated polymer-based SCOSCs and offers valuable guidance for the design of novel double-cable conjugated polymers. 展开更多
关键词 Single-component organic solar cells Double-cable conjugated polymers Photovoltaic properties Alkyl linkers
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木聚糖酶-甘露聚糖酶融合酶基因Linker优化及其在猪肾pK15细胞中共表达 被引量:5
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作者 张献伟 张冠冠 +8 位作者 吴珍芳 孟繁明 刘德武 张茂 许卫华 郑恩琴 贺晓燕 李真 李紫聪 《中国农业科学》 CAS CSCD 北大核心 2013年第22期4774-4783,共10页
[目的]构建木聚糖酶(XynB)与甘露聚糖酶(ManA)的融合酶基因,使其能在哺乳动物表达系统中表达并分泌兼有木聚糖酶和甘露聚糖酶活性的双功能融合酶。[方法]利用基因融合技术(SOE—PCR),把11条Linker融合到木聚糖酶(XynB)和甘露... [目的]构建木聚糖酶(XynB)与甘露聚糖酶(ManA)的融合酶基因,使其能在哺乳动物表达系统中表达并分泌兼有木聚糖酶和甘露聚糖酶活性的双功能融合酶。[方法]利用基因融合技术(SOE—PCR),把11条Linker融合到木聚糖酶(XynB)和甘露聚糖酶(ManA)基因间,构建真核表达载体,经转染猪肾细胞(pKl5)收集细胞培养液,用DNS法测定其酶活并进行酶学分析。[结果]经表达分析12条不同Linker构建的融合酶与亲本酶酶活,发现用Linkera3、pS3和具有“自我剪切”能力T2A构建的融合酶在木聚糖酶和甘露聚糖酶活性都显著高于两亲本酶。融合酶XynB-a3-ManA的木聚糖酶和甘露聚糖酶酶活比亲本酶XynB和ManA分别提高了54.06%和104.40%;该融合酶在酸性环境3.0—7.0起作用,对pH3.08.0具有一定的耐受力。[结论]首次获得可在哺乳动物系统表达分泌的增强型双功能XynB—ManA融合酶。 展开更多
关键词 木聚糖酶 甘露聚糖酶 linker 融合酶 酶活测定
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利用SFX Citaion Linker实现电子图书的跨平台应用 被引量:5
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作者 窦天芳 张喜来 +1 位作者 张成昱 姜爱蓉 《现代图书情报技术》 CSSCI 北大核心 2008年第9期83-86,共4页
阐述清华大学实现电子图书跨平台应用的前期准备工作,介绍SFX Citation Linker的工作机制,提出利用商业系统的标准接口实现电子图书跨平台应用的具体思路和方案。最后,从如何挖掘商业系统的深层功能实现图书馆主动服务的角度给出建议。
关键词 SFX CITATION linker 电子图书 跨平台 主动服务
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有linker插入的靶向核糖核酸酶抑制乙肝病毒复制 被引量:2
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作者 宫卫东 刘军 +3 位作者 丁劲 赵亚 李英辉 薛采芳 《第四军医大学学报》 北大核心 2003年第21期1956-1959,共4页
目的 :构建人嗜酸性粒细胞来源的神经毒素(hEDN )与乙肝病毒核心蛋白 (HBVc)之间插入有linker(Gly4Ser) 3 的融合真核表达载体 (该融合蛋白即为靶向核糖核酸酶 ) ,以优化分子折叠 ,并将其应用于抑制HBV复制的体外研究 .方法 :以第四军... 目的 :构建人嗜酸性粒细胞来源的神经毒素(hEDN )与乙肝病毒核心蛋白 (HBVc)之间插入有linker(Gly4Ser) 3 的融合真核表达载体 (该融合蛋白即为靶向核糖核酸酶 ) ,以优化分子折叠 ,并将其应用于抑制HBV复制的体外研究 .方法 :以第四军医大学病原生物学教研室已构建的pcDNA3.1 (- ) /TR为基础 .首先合成linker序列 ,经过退火形成双链并克隆入 pcDNA3.1 (- ) /TR生成pcDNA3.1 (- ) /HBc linker质粒 ;随后hEDN片段经PCR扩增后克隆入 pcDNA3.1 (- ) /HBc linker质粒生成 pcDNA3.1 (- ) /TNL质粒 ,其中效应分子 (hEDN)与靶向分子 (HBVc)之间为linker序列 .应用间接免疫荧光法检测 pcDNA3.1 (- ) /TNL在细胞的表达 ,并应用放免法测定其抗HBV活性 .同时 ,应用MTT比色法检测细胞的代谢活性 .结果 :成功构建了有linker (Gly4Ser) 3 介入的hEDN和HBVc真核融合表达载体 ;并在HepG2 .2 .1 5细胞中得到有效表达 .转染质粒 pcDNA3.1(- ) /TNL与 pcDNA3.1 (- ) /TR相比可明显地降低HepG2 .2 .1 5细胞上清中HBsAg的含量 (P <0 .0 5 ) .MTT比色分析表明细胞代谢活性未受到影响 (P >0 .0 5 ) .结论 展开更多
关键词 乙肝病毒 linker 核衣壳导向的病毒灭活 靶向核糖核酸酶 抑制效应
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cTnI-linker-TnC融合蛋白基因的构建、表达及鉴定 被引量:2
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作者 张明明 洪理泉 +1 位作者 卢仁泉 郑佐娅 《现代检验医学杂志》 CAS 2007年第1期10-13,共4页
目的重组及表达人cTnI-linker-TnC融合蛋白。方法应用PCR技术从人心脏cDNA文库分别扩增出cTnI和TnC基因,通过引物设计在两者之间加上19个中性氨基酸残基TS-(G4S)3-AC的linker编码序列,克隆PCR产物,并构建pET28a-cTnI-linker-TnC表达质粒... 目的重组及表达人cTnI-linker-TnC融合蛋白。方法应用PCR技术从人心脏cDNA文库分别扩增出cTnI和TnC基因,通过引物设计在两者之间加上19个中性氨基酸残基TS-(G4S)3-AC的linker编码序列,克隆PCR产物,并构建pET28a-cTnI-linker-TnC表达质粒,转化入大肠杆菌表达菌株BL21(DE3)中,用异丙基硫代-β-D-半乳糖苷(IPTG)诱导目的蛋白的表达,NTA树脂亲和层析纯化后检测其纯度和免疫反应性。结果成功构建了cTnI-linker-TnC融合蛋白的基因,并在大肠杆菌中实现可溶性高表达,在摇瓶中的表达量为21 mg/L,经一步NTA树脂亲和层析纯化,获得条带单一的目的蛋白,采用进口全自动免疫检测系统鉴定证实,目的蛋白有较高的免疫反应性。结论采用原核表达方法可以获得具有高纯度和高免疫反应活性的cTnI-linker-TnC融合蛋白。 展开更多
关键词 cTnI—linker—TnC 融合蛋白 表达
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IL-6D_(24)- Linker-PE40重组毒素的分子生物学特性预测 被引量:1
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作者 郑黎燕 奚永志 +4 位作者 孔繁华 陈兴国 金荔 屠敏 刘楠 《中国免疫学杂志》 CAS CSCD 北大核心 1999年第8期339-341,共3页
目的:探讨重组毒素IL6D24LinkerPE40融合蛋白及其接头设计的合理性。方法:应用核酸和蛋白质序列分析软件系统GOLDKEY软件,对IL6D24LinkerPE40重组毒素融合蛋白的柔性、抗原性... 目的:探讨重组毒素IL6D24LinkerPE40融合蛋白及其接头设计的合理性。方法:应用核酸和蛋白质序列分析软件系统GOLDKEY软件,对IL6D24LinkerPE40重组毒素融合蛋白的柔性、抗原性、亲水性、表位等分子生物学特性进行计算机模拟预测,并做Westernblot分析进行验证。结果:IL6D24LinkerPE40分别保留了IL6D24和PE40的表位特征,没有新的抗原表位出现,而且柔性接头处具有极低的抗原性。Westernblot分析结果表明融合蛋白能分别与IL6和PEA的单克隆抗体发生特异结合,这说明融合蛋白保留了IL6和PE40的空间结构特征。 展开更多
关键词 linker PE40 重组毒素 分子结构预测
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Assembly fabrication of linkers on glass surface and their effect on DNA synthesis and hybridization
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作者 沈佳尧 肖鹏峰 +3 位作者 侯鹏 祭美菊 孙啸 何农跃 《Journal of Southeast University(English Edition)》 EI CAS 2003年第2期149-154,共6页
Linkers were assembled on a glass surface based on the hydrolysis and condensation of 3-glycidoxy ̄propyltrimethoxysilane (GPS). After the assembly of GPS, four approaches were tried to open the ending epoxide group o... Linkers were assembled on a glass surface based on the hydrolysis and condensation of 3-glycidoxy ̄propyltrimethoxysilane (GPS). After the assembly of GPS, four approaches were tried to open the ending epoxide group of GPS or to further elongate the linkers. The effect of these approaches on DNA in situ synthesis and hybridization was investigated. For the spacing of the synthesis initiation sites, the wettability of the support and the length of the linking group that attaches the initiation site to the surface have direct influences on the yield of coupling reactions and the subsequent hybridization events. X-ray photoelectron spectroscopy (XPS) and mean contact angles of deionized water of the above slides were measured to assess the linker's characteristics in each procedure. It was proved that the glass slides were successfully modified and became excellent supports for the oligonucleotides synthesis. In addition, it proved best for the in situ oligonucleotides synthesis that a glass slide was in turn treated with ethylenediamine, glutaradehyde, ethanolamine and sodium borohydride solution at ambient temperature after silanized with GPS. 展开更多
关键词 linker assembly fabrication in situ oligonucleotide synthesis HYBRIDIZATION
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可溶性sCD80-Linker-sCD40L融合蛋白诱导非特异性抗肿瘤免疫
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作者 徐冬 魏枫 +2 位作者 付晓达 于津浦 任秀宝 《中国肿瘤生物治疗杂志》 CAS CSCD 2007年第5期471-476,共6页
目的:观察可溶性sCD80-Linker-sCD40L融合蛋白对体外非特异性抗肿瘤免疫的影响。方法:以含sCD80-Lin- ker-sCD40L、sCD80、sCD40L编码基因的重组腺病毒载体或对照病毒载体转染人卵巢癌细胞SKOV3,ELISA检测上清中可溶性sCD80-Linker-sCD... 目的:观察可溶性sCD80-Linker-sCD40L融合蛋白对体外非特异性抗肿瘤免疫的影响。方法:以含sCD80-Lin- ker-sCD40L、sCD80、sCD40L编码基因的重组腺病毒载体或对照病毒载体转染人卵巢癌细胞SKOV3,ELISA检测上清中可溶性sCD80-Linker-sCD40L融合蛋白及sCD80、sCD40L蛋白的表达。由卵巢癌患者外周血单个核细胞培养树突状细胞(dendritic cells,DCs),将DCs和自体T细胞共同培养并加入不同上清诱导48h;用异基因混合淋巴细胞反应(mixed lymphocyte reaction,MLR)检测诱导的DCs刺激淋巴细胞增殖能力;以诱导的T细胞为效应细胞,SKOV3细胞或K562细胞为非特异性靶细胞,通过乳酸脱氢酶(lactate dehydrogemse,LDH)释放实验检测杀伤活性。结果:ELISA检测证实病毒转染SKOV3细胞上清中sCD80-Linker-sCD40L融合蛋白、sCD80蛋白和sCD40L蛋白表达量分别为2.791 ng/ml、1.956 ng/ml和1.407 ng/ml。MLR证实,与对照相比,融合蛋白上清诱导的DCs[刺激-反应细胞比例1:100,(0.382±0.053)vs(0.167±0.028),P<0.01;刺激-反应细胞比例1:50,(0.636±0.164)vs(0.311±0.067),P<0.01;刺激-反应细胞比例1:10,(1.245±0.271)vs(0.423±0.156),P<0.01]和sCD40L上清诱导的DCs[刺激-反应细胞比例1:100,(0.341±0.062)vs(0.167±0.028),P<0.01;刺激-反应细胞比例1:50,(0.567±0.106)vs(0.311±0.067),P<0.01;刺激-反应细胞比例1:10,(1.098±0.263)vs(0.423±0.156),P<0.01]刺激同种异基因混合淋巴细胞增殖能力明显增强。LDH释放实验提示,sCD40L上清诱导的T淋巴细胞对靶细胞非特异性杀伤活性显著强于对照SKOV3组[(42.28±6.27)vs(22.49±3.56),P<0.01]和K562组(47.94±6.72)vs (26.33±2.89),P<0.01],而融合蛋白上清诱导的T淋巴细胞对靶细胞非特异性杀伤活性显著强于sCD40L上清诱导的T淋巴细胞[sKOV3组为(58.97±8.92)vs(42.28±6.27),P<0.05;K562组为(66.55±9.43)vs(47.94±6.72),P<0.05]。结论:可溶性sCD80-Linker-sCD40L融合蛋白能有效诱导体外非特异性抗肿瘤免疫,对肿瘤免疫治疗具有潜在应用价值。 展开更多
关键词 CD80 CD40L sCD80-linker—sCD40L 融合蛋白 树突状细胞 卵巢癌细胞 抗肿瘤免疫
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固相合成蛋白所需生物大分子Linker-Met和fma-tRNA的构建(英文)
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作者 侯文 刘传法 《南开大学学报(自然科学版)》 CAS CSCD 北大核心 2017年第4期14-27,共14页
无细胞表达蛋白质体系近年来恢复活力,以满足日益增长的蛋白合成需求.与传统的细胞重组表达蛋白的方法相比,无细胞体系具有许多优点,包括更快的速度、更高的通量和更大的灵活性.此外,它提供了独特的可能性,如合理的DNA模板设计,毒素蛋... 无细胞表达蛋白质体系近年来恢复活力,以满足日益增长的蛋白合成需求.与传统的细胞重组表达蛋白的方法相比,无细胞体系具有许多优点,包括更快的速度、更高的通量和更大的灵活性.此外,它提供了独特的可能性,如合理的DNA模板设计,毒素蛋白的表达,多个多肽链的联合表达,高效的定位标记等等.在本文中,建立了一个linker-tRNA的方法利用无细胞体系固相合成蛋白质.首先,需要将第1个氨基酸连接上起始tRNA通过linker连接到固相表面.为了提高固相合成蛋白质的效率,利用含有琥珀终止子(CUA)为反密码子的linker-tRNA与含互补琥珀子(UAG)为起始密码子的DNA模板而不是正常的起始密码子(密码子AUG).已经成功地获得了纯化的fma-tRNA(-CA)和linker-Met了.此方法将为蛋白质芯片的开发提供新途径. 展开更多
关键词 蛋白质固相合成 fma-tRNA的合成 linker-Met的合成
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cTnI(28-110aa)-linker-TnC融合蛋白基因的构建、表达及鉴定
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作者 洪理泉 郑佐娅 赵卫国 《中国微循环》 北大核心 2005年第6期380-383,共4页
目的构建及表达人cTnI(28-110 aa)-linker-TnC融合蛋白。方法应用PCR技术从人心脏cDNA文库中扩增出cTnI(28-110 aa)和TnC基因,通过引物设计在两者之间加上linker序列即编码19个中性氨基酸残基TS-(G4S)3-AC的序列,克隆PCR产物,并构建成pE... 目的构建及表达人cTnI(28-110 aa)-linker-TnC融合蛋白。方法应用PCR技术从人心脏cDNA文库中扩增出cTnI(28-110 aa)和TnC基因,通过引物设计在两者之间加上linker序列即编码19个中性氨基酸残基TS-(G4S)3-AC的序列,克隆PCR产物,并构建成pET28 a-cTnI(28-110 aa)-lin-ker-TnC表达质粒,转化入大肠杆菌表达菌株BL21(DE3),用异丙基硫代-β-D-半乳糖苷(IPTG)诱导目的蛋白的表达,NTA树脂亲和层析纯化后检测其纯度和免疫反应性。结果成功构建了cTnI(28-110 aa)-linker-TnC融合蛋白的基因,并在大肠杆菌中实现可溶性高表达,在摇瓶中的表达量为20 mg/L,经一步NTA树脂亲和层析纯化,获得条带单一的目的蛋白,采用进口全自动免疫检测系统鉴定证实,目的蛋白有较高的免疫反应性。结论采用原核表达方法获得了具有高纯度和高免疫反应活性的cTnI(28-110 aa)-linker-TnC融合蛋白。 展开更多
关键词 cTnI(28-110aa)-linker-TnC 融合蛋白 表达
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绿色木霉TvALP-linker-TvRBL融合基因的构建与原核表达 被引量:4
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作者 徐杨玉 温世杰 +2 位作者 李海芬 李玲 梁炫强 《生物技术通报》 CAS CSCD 北大核心 2015年第1期131-137,共7页
在Tv ALP基因和Tv RBL基因间加入一段柔性链接头(linker)基因序列,构建融合基因。生物信息学分析表明该融合基因含有一段信号肽序列。利用RT-PCR技术从绿色木霉菌丝体总RNA中扩增出Tv RBL基因、Tv ALP基因和去除信号肽的ΔTvALP基因,分... 在Tv ALP基因和Tv RBL基因间加入一段柔性链接头(linker)基因序列,构建融合基因。生物信息学分析表明该融合基因含有一段信号肽序列。利用RT-PCR技术从绿色木霉菌丝体总RNA中扩增出Tv RBL基因、Tv ALP基因和去除信号肽的ΔTvALP基因,分别克隆到原核表达载体p ET30a,构建重组质粒p ET30a-Tv ALP-linker-Tv RBL和p ET30a-ΔTv ALP-linker-Tv RBL,转化大肠杆菌BL21(DE3)p Lys S,经异丙基硫代-β-D-半乳糖苷(IPTG)诱导表达。SDS-PAGE电泳分析结果表明,去除信号肽的表达质粒p ET30a-ΔTv ALP-linker-Tv RBL在大肠杆菌BL21(DE3)p Lys S中获得了表达,在60 k D处有一条蛋白质特异条带,与预测的目的产物蛋白条带大小一致。 展开更多
关键词 绿色木霉 TvALP-linker-TvRBL融合基因 信号肽 原核表达
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cTnI-linker-TnC融合蛋白的原核表达及鉴定 被引量:1
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作者 龚隆财 罗镇明 +3 位作者 杨雁青 王振宇 向军俭 王宏 《中国生物工程杂志》 CAS CSCD 北大核心 2015年第4期48-53,共6页
从PUBMED中查找人心脏cTnI和cTnC的cDNA序列,在两者之间加上15个中性氨基酸残基(G4S)3的linker编码序列,并分别构建原核表达质粒pET32a-cTnI-linker-TnC和pET28a-cTnI-linker-TnC,转化入大肠杆菌表达菌株BL21(DE3)中,用异丙基硫代-β-D... 从PUBMED中查找人心脏cTnI和cTnC的cDNA序列,在两者之间加上15个中性氨基酸残基(G4S)3的linker编码序列,并分别构建原核表达质粒pET32a-cTnI-linker-TnC和pET28a-cTnI-linker-TnC,转化入大肠杆菌表达菌株BL21(DE3)中,用异丙基硫代-β-D-半乳糖苷(IPTG)诱导其表达。通过SDS-PAGE电泳和间接ELISA对目的蛋白进行初步鉴定,并优化表达条件以实现目的蛋白的可溶性高表达。用标准抗cTnI单克隆抗体和标准抗cTnC单克隆抗体对目的蛋白进行Western blot鉴定和双抗体夹心ELISA鉴定,同时用八位心肌梗塞患者血清做间接ELISA鉴定。结果表明,cTnI-linker-TnC融合蛋白在表达载体pET32a中实现了可溶性高表达,最佳表达条件为28℃、0.4mmol/L IPTG、诱导表达5h;且融合蛋白具有较好的免疫原性,保存了cTnI和cTnC单体的天然结构,有望成为天然cTnI-TnC复合物的替代物,为下一步制备高特异性的抗cTnI-TnC复合物的单克隆抗体奠定了基础。 展开更多
关键词 cTnI-linker-TnC 融合蛋白 原核表达 双抗体夹心ELISA
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Randomized Terminal Linker-dependent PCR: A Versatile and Sensitive Method for Detection of DNA Damage 被引量:10
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作者 ZHANGZHI-WEI HENGZHENG-CHANG 《Biomedical and Environmental Sciences》 SCIE CAS CSCD 2002年第3期203-208,共6页
Objective To design and develop a novel, sensitive and versatile method for in vivo foot printing and studies of DNA damage, such as DNA adducts and strand breaks. Methods Starting with mammalian genomic DNA, singl... Objective To design and develop a novel, sensitive and versatile method for in vivo foot printing and studies of DNA damage, such as DNA adducts and strand breaks. Methods Starting with mammalian genomic DNA, single-stranded products were made by repeated primer extension, these products were ligated to a double-stranded linker having a randomized 3 overhang, and used for PCR. DNA breaks in p53 gene produced by restriction endonuclease AfaI were detected by using this new method followed by Southern hybridization with DIG-labeled probe. Results This randomized terminal linker-dependent PCR (RDPCR) method could generate band signals many-fold stronger than conventional ligation-mediated PCR (LMPCR), and it was more rapid, convenient and accurate than the terminal transferase-dependent PCR (TDPCR). Conclusion DNA strand breakage can be detected sensitively in the gene level by RDPCR. Any lesion that blocks primer extension should be detectable. 展开更多
关键词 Randomized terminal linker-dependent PCR LMPCR p53 gene DNA damage
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连接肽对融合蛋白ELP[I]_(30)-linker-eGFP相变的影响 被引量:1
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作者 张立超 李军明 +2 位作者 葛高顺 孔令岭 胡学军 《生命科学研究》 CAS CSCD 北大核心 2013年第5期401-405,共5页
研究G4S和Poly N连接肽对融合蛋白ELP[I]30-linker-eGFP相变的影响.将编码两种不同连接肽G4S和Poly N的绿色荧光蛋白(enhanced green fluorescent protein,eGFP)基因克隆到pET28-ELP[I]30表达载体中,在宿主菌E.coli BLR(DE3)中经IPTG诱... 研究G4S和Poly N连接肽对融合蛋白ELP[I]30-linker-eGFP相变的影响.将编码两种不同连接肽G4S和Poly N的绿色荧光蛋白(enhanced green fluorescent protein,eGFP)基因克隆到pET28-ELP[I]30表达载体中,在宿主菌E.coli BLR(DE3)中经IPTG诱导表达ELP[I]30-linker-eGFP,通过可逆相变循环(inverse transition cycling,ITC)及镍柱亲和层析纯化ELP[I]30-linker-eGFP蛋白.结果显示,成功构建、表达具有活性的两种连接肽的融合蛋白ELP[I]30-linker-eGFP,连接肽G4S使融合蛋白产生不可逆相变,而Poly N不影响融合蛋白可逆相变,该研究对类弹性蛋白标签的应用具有指导意义. 展开更多
关键词 连接肽 融合蛋白 纯化标签 类弹性蛋白 绿色荧光蛋白
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Imidazole linker‐induced covalent triazine framework-ZIF hybrids for confined hollow carbon super‐heterostructures toward a long‐life supercapacitor 被引量:3
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作者 Madagonda MVadiyar Ji‐Young Kim +1 位作者 Jee‐Hwan Bae Kyung‐Wan Nam 《Carbon Energy》 SCIE EI CAS CSCD 2023年第10期25-39,共15页
Carbon super-heterostructures with high nitrogen contents from the covalent hybrid precursors of covalent triazine frameworks(CTFs)and zeolitic imidazolic frameworks(ZIFs)are scarcely explored because of CTF's ord... Carbon super-heterostructures with high nitrogen contents from the covalent hybrid precursors of covalent triazine frameworks(CTFs)and zeolitic imidazolic frameworks(ZIFs)are scarcely explored because of CTF's ordered structure and toxic superacid that dissolves or destabilizes the metal nodes.To solve this problem,herein,we report a straightforward two-step pathway for the covalent hybridization of disordered CTF(d–CTF)–ZIF composites via preincorporation of an imidazole(IM)linker into ordered CTFs,followed by the imidazole-site-specific covalent growth of ZIFs.Direct carbonization of these synthesized d–CTF−IM−ZIF hybrids results in unique hollow carbon super-heterostructures with ultrahigh nitrogen content(>18.6%),high specific surface area(1663m^(2)g^(−1)),and beneficial trace metal(Co/Zn NPs)contents for promoting the redox pseudocapacitance.As proof of concept,the obtained carbon super-heterostructure(Co–Zn–NC_(SNH)–800)is used as a positive electrode in an asymmetric supercapacitor,demonstrating a remarkable energy density of 61Wh kg^(−1)and extraordinary cyclic stability of 97%retention after 30,000 cycles at the cell level.Our presynthetic modifications of CTF and their covalent hybridization with ZIF crystals pave the way toward new design strategies for synthesizing functional porous carbon materials for promising energy applications. 展开更多
关键词 asymmetric supercapacitor defective covalent triazine frameworks hollow carbon superheterostructures linker defect
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Multifunctionality of the linker histones: an emerging role for protein-protein interactions 被引量:2
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作者 Steven J McBryant Xu Lu Jeffrey C Hansen 《Cell Research》 SCIE CAS CSCD 2010年第5期519-528,共10页
Linker histones, e.g., H1, are best known for their ability to bind to nucleosomes and stabilize both nucleosome structure and condensed higher-order chromatin structures. However, over the years many investigators ha... Linker histones, e.g., H1, are best known for their ability to bind to nucleosomes and stabilize both nucleosome structure and condensed higher-order chromatin structures. However, over the years many investigators have reported specific interactions between linker histones and proteins involved in important cellular processes. The purpose of this review is to highlight evidence indicating an important alternative mode of action for H1, namely protein-protein interactions. We first review key aspects of the traditional view of linker histone action, including the importance of the H1 C-terminal domain. We then discuss the current state of knowledge of linker histone interactions with other proteins, and, where possible, highlight the mechanism of linker histone-mediated protein-protein interactions. Taken together, the data suggest a combinatorial role for the linker histones, functioning both as primary chromatin architectural proteins and simultaneously as recruitment hubs for proteins involved in accessing and modifying the chromatin fiber. 展开更多
关键词 CHROMATIN linker histone higher-order structure NUCLEOSOMES
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Effects of linker amino acids on the potency and selectivity of dimeric antimicrobial peptides 被引量:2
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作者 Ming Kai Wei Zhang +8 位作者 Huan Xie Liwei Liu Sujie Huang Xiao Li Zhengzheng Zhang Yuyang Liu Bangzhi Zhang Jingjing Song Rui Wang 《Chinese Chemical Letters》 SCIE CAS CSCD 2018年第7期1163-1166,共4页
Dimerization is an effective strategy for designing antimicrobial peptides that combine the advantages of different native peptides. In this study, we explored the effects of different linker amino acids, including le... Dimerization is an effective strategy for designing antimicrobial peptides that combine the advantages of different native peptides. In this study, we explored the effects of different linker amino acids, including leucine, proline and aminocaproic acid, on the anticancer, antimicrobial and hemolytic activities of the heteromeric antimicrobial peptides AM-1, AM-2, and AM-3. Proline and aminocaproic acid are ideal linkers for increasing the potency and selectivity of heteromeric antimicrobial peptides. The results of MD simulations provided a rationalization for this observation. Both AM-2, which had a proline linker,and AM-3, which had an aminocaproic acid linker, adopted a compact conformation in water and a bent conformation in membranes. This change in the flexible structures of AM-2 and AM-3 could have resulted in decreased binding of these peptides to zwitterionic lipid bilayers and increased damage to mixed lipid bilayers containing acidic phospholipids. In short, these findings obtained via assessing the effects of linker amino acids will contribute to the design of ideal heteromeric antimicrobial peptides with high selectivity and potency. 展开更多
关键词 linker amino acid Flexibility Heteromeric antimicrobial peptides Potency and selectivity
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