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Car9-tag介导的重组ω-转氨酶的高效表达和纯化 被引量:1
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作者 张洪起 李姣姣 +1 位作者 周丽亚 高静 《化学研究与应用》 CAS CSCD 北大核心 2018年第8期1252-1257,共6页
对来源于紫色色杆菌的ω-转氨酶基因(Genebank:JC032687.1)进行了生物信息学分析,其理论分子量为51213.42,是一种含有多个磷酸功能位点的亲水性蛋白质;利用p ET-28a(+)为载体在宿主细胞E.coli.Rosetta(DE3)中重组表达了融合有Car9标签(C... 对来源于紫色色杆菌的ω-转氨酶基因(Genebank:JC032687.1)进行了生物信息学分析,其理论分子量为51213.42,是一种含有多个磷酸功能位点的亲水性蛋白质;利用p ET-28a(+)为载体在宿主细胞E.coli.Rosetta(DE3)中重组表达了融合有Car9标签(Car9-tag)的ω-转氨酶;考察了诱导条件等对ω-转氨酶表达量的影响,摇瓶培养2 h后加入终浓度为1.0 m M的异丙基-β-D-硫代吡喃半乳糖苷(IPTG)诱导10 h后可获得高表达量的融合ω-转氨酶,其比酶活可达1 547.16 U·g-1;采用了二氧化硅亲和吸附Car9-tagω-转氨酶对ω-转氨酶进行了分离纯化。 展开更多
关键词 ω-转氨酶 car9-tag 基因重组 高效表达
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基于MATLAB与ADAMAS的磁流变减振器整车联合仿真 被引量:1
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作者 王祺明 祝辉 《客车技术》 2008年第3期13-18,21,共7页
在对磁流变减振器工作原理分析的基础上.针对目前理论研究中对阻尼力实施控制和没有建立精确的整车模型而带来的仿真与实际情况差别较大的问题,在SIMULINK中建立了磁流变减振器的数学模型。结合其工作原理设计了以车身加速度和簧载质... 在对磁流变减振器工作原理分析的基础上.针对目前理论研究中对阻尼力实施控制和没有建立精确的整车模型而带来的仿真与实际情况差别较大的问题,在SIMULINK中建立了磁流变减振器的数学模型。结合其工作原理设计了以车身加速度和簧载质量与非簧载质量相对速度为输入,控制电流为输出的模糊控制器。在ADAMS/CAR中建立了比较准确的基于磁流变减振器的半主动悬架及整车多刚体动力学模型,并进行了联合仿真。结果表明了磁流变减振器能够有效提高车辆的平顺性与舒适性,同时也为加快磁流变减振器研发提供了理论依据。 展开更多
关键词 SIMULINK ADAM9CAR 磁流变减振器 多刚体动力学 联合仿真 平顺性
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LOWER TEMPERATURE 1 Enhances ABA Responses and Plant Drought Tolerance by Modulating the Stability and Localization of C2-Domain ABA-Related Proteins in Arabidopsis 被引量:3
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作者 Tao Qin Qiuzhen Tian +1 位作者 Guifeng Wang Liming Xiong 《Molecular Plant》 SCIE CAS CSCD 2019年第9期1243-1258,共16页
Plasma membrane-associated abscisic acid(ABA)signal transduction is an integral part of ABA signaling.The C2-domain ABA-related(CAR)proteins play important roles in the recruitment of ABA receptors to the plasma membr... Plasma membrane-associated abscisic acid(ABA)signal transduction is an integral part of ABA signaling.The C2-domain ABA-related(CAR)proteins play important roles in the recruitment of ABA receptors to the plasma membrane to facilitate ABA signaling.However,how CAR proteins are regulated remains unclear.In this study,we conducted a genetic screen for mutants with altered leaf transpiration and identified an uncharacterized protein,LOWER TEMPERATURE 1(LOT1),which regulates the dynamic localization and stability of CAR proteins.The lotimutant had a lower leaf temperature as compared with the wild type due to higher transpiration.We found that LOT1 physically interacts with CAR9,and ABA reduces LOT1-CAR9 interaction in the nucleus,likely via Ca^2+,resulting in increased localization of CAR9 to the plasma membrane.We further found that the stability of CAR9 is affected by LOT1 less CAR9 proteins were accumulated and more were ubiquitinated in lot1.While the lot1 car9 and/of f car9 mutants were hyposerisitive to ABA,the hyposensitive phenotype of loticould be rescued by CAR9 overexpression.Collectively,our study reveals that LOT1 regulates plant tolerance to drought stress by affecting ABA signaling through regulating the stability and dynamic localization of CAR9. 展开更多
关键词 LOT1 car9 DROUGHT tolerance ABA signaling STOMATA TRANSPIRATION
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