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61.
Thi Kieu Oanh Nguyen Thi Luong Vu Minh Quan Nguyen Huynh Kim Khanh Ta Kyoung Sun Park Soo Hyeon Kim Chong Jai Kim Yeon Jin Jang Han Choe 《International journal of molecular sciences》2021,22(10)
Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a member of the colony-stimulating factor (CSF) family, which functions to enhance the proliferation and differentiation of hematopoietic stem cells and other hematopoietic lineages such as neutrophils, dendritic cells, or macrophages. These proteins have thus generated considerable interest in clinical therapy research. A current obstacle to the prokaryotic production of human GM-CSF (hGM-CSF) is its low solubility when overexpressed and subsequent complex refolding processes. In our present study, the solubility of hGM-CSF was examined when combined with three N-terminal fusion tags in five E. coli strains at three different expression temperatures. In the five E. coli strains BL21 (DE3), ClearColi BL21 (DE3), LOBSTR, SHuffle T7 and Origami2 (DE3), the hexahistidine-tagged hGM-CSF showed the best expression but was insoluble in all cases at each examined temperature. Tagging with the maltose-binding protein (MBP) and the b′a′ domain of protein disulfide isomerase (PDIb′a′) greatly improved the soluble overexpression of hGM-CSF at 30 °C and 18 °C. The solubility was not improved using the Origami2 (DE3) and SHuffle T7 strains that have been engineered for disulfide bond formation. Two conventional chromatographic steps were used to purify hGM-CSF from the overexpressed PDIb′a′-hGM-CSF produced in ClearColi BL21 (DE3). In the experiment, 0.65 mg of hGM-CSF was isolated from a 0.5 L flask culture of these E. coli and showed a 98% purity by SDS-PAGE analysis and silver staining. The bioactivity of this purified hGM-CSF was measured at an EC50 of 16.4 ± 2 pM by a CCK8 assay in TF-1 human erythroleukemia cells. 相似文献
62.
Sean R. Tachibana Longteng Tang Liangdong Zhu Yuka Takeda Keiji Fushimi Yoshibumi Ueda Takahiro Nakajima Yuto Kuwasaki Moritoshi Sato Rei Narikawa Chong Fang 《International journal of molecular sciences》2021,22(10)
Cyanobacteriochromes (CBCRs) are promising optogenetic tools for their diverse absorption properties with a single compact cofactor-binding domain. We previously uncovered the ultrafast reversible photoswitching dynamics of a red/green photoreceptor AnPixJg2, which binds phycocyanobilin (PCB) that is unavailable in mammalian cells. Biliverdin (BV) is a mammalian cofactor with a similar structure to PCB but exhibits redder absorption. To improve the AnPixJg2 feasibility in mammalian applications, AnPixJg2_BV4 with only four mutations has been engineered to incorporate BV. Herein, we implemented femtosecond transient absorption (fs-TA) and ground state femtosecond stimulated Raman spectroscopy (GS-FSRS) to uncover transient electronic dynamics on molecular time scales and key structural motions responsible for the photoconversion of AnPixJg2_BV4 with PCB (Bpcb) and BV (Bbv) cofactors in comparison with the parent AnPixJg2 (Apcb). Bpcb adopts the same photoconversion scheme as Apcb, while BV4 mutations create a less bulky environment around the cofactor D ring that promotes a faster twist. The engineered Bbv employs a reversible clockwise/counterclockwise photoswitching that requires a two-step twist on ~5 and 35 picosecond (ps) time scales. The primary forward Pfr → Po transition displays equal amplitude weights between the two processes before reaching a conical intersection. In contrast, the primary reverse Po → Pfr transition shows a 2:1 weight ratio of the ~35 ps over 5 ps component, implying notable changes to the D-ring-twisting pathway. Moreover, we performed pre-resonance GS-FSRS and quantum calculations to identify the Bbv vibrational marker bands at ~659,797, and 1225 cm−1. These modes reveal a stronger H-bonding network around the BV cofactor A ring with BV4 mutations, corroborating the D-ring-dominant reversible photoswitching pathway in the excited state. Implementation of BV4 mutations in other PCB-binding GAF domains like AnPixJg4, AM1_1870g3, and NpF2164g5 could promote similar efficient reversible photoswitching for more directional bioimaging and optogenetic applications, and inspire other bioengineering advances. 相似文献
63.
摘要:日本福岛核电站泄漏事故暴露了锆合金易与水蒸气反应造成爆炸的缺陷,又将人们的视线拉回了各项性能较为均衡的FeCrAl合金。FeCrAl合金较锆合金具有更好的耐腐蚀性能,更能满足核反应堆高温水蒸气环境下的工作条件。为进一步保障核反应堆安全性,各国学者对FeCrAl合金的性能进行了大量研究。基于此,综述了FeCrAl合金腐蚀机制及其在高温蒸汽和高温水环境中的腐蚀行为;探讨了合金元素(稀土、Ti、Zr、Nb和Mo)、变形和热处理对合金力学性能的影响;总结了FeCrAl合金辐照性能和焊接性能的研究进展。最后指出了核级FeCrAl合金在研究中存在的不足及未来重点发展方向,为今后其他学者在该领域的研究提供参考。 相似文献
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通过分析高强度低合金钢焊缝熔敷金属常见显微组织对其力学性能的影响,确定X80管线钢用埋弧焊丝熔敷金属组织应以大量针状铁素体(AF)和少量粒状贝氏体(GB)的复合组织.从相变动力学原理出发,结合针状铁素体(AF)非自发形核机制和微合金组织韧化理论,选择Mn-Ni-Mo-Ti-B合金系进行X80管线钢匹配焊丝的试制.结果表明,合理选择和控制合金元素,可以获得理想的焊缝熔敷金属组织和强韧性以及低温韧性要求,试制的1号焊丝能够满足X80管线钢的使用要求. 相似文献
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1 INTRODUCTIONComparedwithamixer settlerorpulsedcolumnusedinthenuclearfuelreprocessingextraction pro cesses,ashort residence timecentrifugalextractorcatchesresearcher seyesbecauseofthe goodphaseseparationandasmallhold upvolume .Itshowsthefollowingadvantagesinthenuclearindustry[1] :1)Reducingsolventdegradationduetoradiation ;2 )Reducinginventoryofaqueousandorganicphases ,whichgivesrapidstart upandshut down ;3)Com pactdesignthatcontributestothereductionofplantcost .Manydifferenttypesofcen… 相似文献
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结合国家“十五”重点科技攻关计划“城市规划、建设、管理与服务数字化工程”项目,天津水泥工业设计研究院承担了其中的“勘察设计企业信息化应用软件示范应用”专题,并获批立项,签署了专题任务书。两年来,院项目实施小组和软件公司进行了长期、紧密的合作,遵照标准化、系统化和集成化的原则,共同搭建了“工程协同设计管理平台”。在软件功能上,实现了设计企业管理信息系统的主要功能;在软件技术上,采用了目前最流行的软件技术,结合现代项目管理方法,为设计企业提供了组态灵活,方便实用,具有良好扩展性及开放性的软件产品。目前,该系统已经在部分工程项目上全面运行,取得了预期的效果。 相似文献
70.
The mobile wireless market has been attracting many customers. Technically, the paradigm of anytime-anywhere connectivity raises previously unthinkable challenges, including the management of million of mobile customers, their profiles, the profiles-based selective information dissemination, and server-side computing infrastructure design issues to support such a large pool of users automatically and intelligently. In this paper, we propose a data mining technique for discovering frequent behavioral patterns from a collection of trajectories gathered by Global Positioning System. Although the search space for spatiotemporal knowledge is extremely challenging, imposing spatial and temporal constraints on spatiotemporal sequences makes the computation feasible. Specifically, the mined patterns are incorporated with synthetic constraints, namely spatiotemporal sequence length restriction, minimum and maximum timing gap between events, time window of occurrence of the whole pattern, inclusion or exclusion event constraints, and frequent movement patterns predictive of one ore more classes. The algorithm for mining all frequent constrained patterns is named cAllMOP. Moreover, to control the density of pattern regions a clustering algorithm is exploited. The proposed method is efficient and scalable. Its efficiency is better than that of the previous algorithms AllMOP and GSP with respect to the compactness of discovered knowledge, execution time, and memory requirement. 相似文献