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41.
对赤红球菌的组氨酸激酶基因进行密码子优化,将优化后的组氨酸激酶基因(rhks)构建重组表达质粒pGEX-4T-2-rhks。将此质粒导入到大肠杆菌BL21(DE3)中进行异源表达。在25 ℃和1 mmol/L异丙基-β-D-硫代吡喃半乳糖苷诱导条件下,组氨酸激酶融合蛋白(GST-RHK)获得成功表达,并具有催化活性。经谷胱甘肽琼脂糖亲和层析纯化,获得电泳纯的GST-RHK,其中纯化倍数为3.1,得率为19.5%。该蛋白大小约为72.75 kDa,Km、Vmax和Kcat值分别为20.92 μmol/L、0.17 μmol/(L·min)和1.4 min-1。野生型赤红球菌、组氨酸激酶基因增强株sdrhkE和组氨酸激酶基因敲减株sdrhkD在分别含有苯酚、甲苯、氯苯、异辛烷4 种有机溶剂的培养基中培养,菌株sdrhkD的生长情况都优于野生型赤红球菌,菌株sdrhkE的生长情况都低于野生型赤红球菌。本研究为进一步揭示赤红球菌SD3中组氨酸激酶涉及的信号转导途径与赤红球菌有机溶剂耐受性的关联机制提供一定参考依据。 相似文献
42.
Lorenzo Zallocco Laura Giusti Maurizio Ronci Andrea Mussini Marco Trerotola Maria Rosa Mazzoni Antonio Lucacchini Laura Sebastiani 《International journal of molecular sciences》2021,22(9)
The autonomic nervous system (ANS) plays a crucial role both in acute and chronic psychological stress eliciting changes in many local and systemic physiological and biochemical processes. Salivary secretion is also regulated by ANS. In this study, we explored salivary proteome changes produced in thirty-eight University students by a test stress, which simulated an oral exam. Students underwent a relaxation phase followed by the stress test during which an electrocardiogram was recorded. To evaluate the effect of an olfactory stimulus, half of the students were exposed to a pleasant odor diffused in the room throughout the whole session. Saliva samples were collected after the relaxation phase (T0) and the stress test (T1). State anxiety was also evaluated at T0 and T1. Salivary proteins were separated by two-dimensional electrophoresis, and patterns at different times were compared. Spots differentially expressed were trypsin digested and identified by mass spectrometry. Western blot analysis was used to validate proteomic results. Anxiety scores and heart rate changes indicated that the fake exam induced anxiety. Significant changes of α-amylase, polymeric immunoglobulin receptor (PIGR), and immunoglobulin α chain (IGHA) secretion were observed after the stress test was performed in the two conditions. Moreover, the presence of pleasant odor reduced the acute social stress affecting salivary proteome changes. Therefore, saliva proteomic analysis was a useful approach to evaluate the rapid responses associated to an acute stress test also highlighting known biomarkers. 相似文献
43.
以转基因水稻中最常用的CaMV35S启动子、NOS终止子、Cry1Ab/Ac基因、HPT基因及SPS水稻内标基因为研究对象,利用5 种不同的荧光信号(FAM、HEX、Taxas Red、Cy5、Cy5.5)进行多重实时聚合酶链式反应(real-time polymerase chain reaction,real-time PCR)检测方法的研究。通过引物组合筛选、反应体系优化、特异性测试、灵敏度测试、适用性测试等一系列实验,建立了5 重real-time PCR方法,灵敏度可达0.032%。此方法具有灵敏度高、结果准确、通量大等优点,可实现水稻中转基因成分的快速、高效检测。 相似文献
44.
45.
《Intermetallics》2015
Raney-type Ni precursor alloys containing 75 at.% Al and doped with 0, 0.75, 1.5 and 3.0 at.% Ti have been produced by a gas atomization process. The resulting powders have been classified by size fraction with subsequent investigation by powder XRD, SEM and EDX analysis. The undoped powders contain, as expected, the phases Ni2Al3, NiAl3 and an Al-eutectic. The Ti-doped powders contain an additional phase with the TiAl3 DO22 crystal structure. However, quantitative analysis of the XRD results indicate a far greater fraction of the TiAl3 phase is present than could be accounted for by a simple mass balance on Ti. This appears to be a (TixNi1−x)Al3 phase in which higher cooling rates favour small x (low Ti-site occupancy by Ti atoms). SEM and EDX analysis reveal that virtually all the available Ti is contained within the TiAl3 phase, with negligible Ti dissolved in either the Ni2Al3 or NiAl3 phases. 相似文献
46.
Zhuang CHENG Jianfeng WANG Matthew Richard COOP Guanlin YE 《Frontiers of Structural and Civil Engineering》2020,14(2):357
The development of a miniature triaxial apparatus is presented. In conjunction with an X-ray micro-tomography (termed as X-ray μCT hereafter) facility and advanced image processing techniques, this apparatus can be used for in situ investigation of the micro-scale mechanical behavior of granular soils under shear. The apparatus allows for triaxial testing of a miniature dry sample with a size of (diameter height). In situ triaxial testing of a 0.4–0.8 mm Leighton Buzzard sand (LBS) under a constant confining pressure of 500 kPa is presented. The evolutions of local porosities (i.e., the porosities of regions associated with individual particles), particle kinematics (i.e., particle translation and particle rotation) of the sample during the shear are quantitatively studied using image processing and analysis techniques. Meanwhile, a novel method is presented to quantify the volumetric strain distribution of the sample based on the results of local porosities and particle tracking. It is found that the sample, with nearly homogenous initial local porosities, starts to exhibit obvious inhomogeneity of local porosities and localization of particle kinematics and volumetric strain around the peak of deviatoric stress. In the post-peak shear stage, large local porosities and volumetric dilation mainly occur in a localized band. The developed triaxial apparatus, in its combined use of X-ray μCT imaging techniques, is a powerful tool to investigate the micro-scale mechanical behavior of granular soils. 相似文献
47.
V.I. Popkov O.V. Almjasheva V.N. Nevedomskiy V.V. Panchuk V.G. Semenov V.V. Gusarov 《Ceramics International》2018,44(17):20906-20912
Formation and structural transformations of yttrium orthoferrite crystals have been studied using X-ray diffractometry, Mössbauer spectroscopy and transmission electron microscopy combined with electron microdiffraction. Said processes have been studied under heat treatment of glycine-nitrate combustion products. There have been identified formations of three structural yttrium orthoferrite modifications – amorphized hexagonal <h1>-YFeO3 (P63cm) and nanocrystalline hexagonal h2-YFeO3 (P63/mmc), as well as nanocrystalline orthorhombic o-YFeO3 (Pbnm), which are selectively formed depending on available three-dimensional confinements. Based on the analysis of changes in the fluid and size composition formulation, it has been proposed mechanism for formation and transformation of YFeO3 nanocrystals, including growth stage of h2-YFeO3 crystals due to amorphized phase of <h1>-YFeO3 up to critical size of about 15?nm and their subsequent transformation into orthorhombic form o-YFeO3. 相似文献
48.
Mingzhi Zhao Jianwei Cao Zhi Wang Guohua Li 《Journal of the European Ceramic Society》2019,39(7):2427-2435
The crystallization phenomena of spinel in CaO-MgO-Al2O3-SiO2-Fe2O3 glass have received much attention due to the particular role in preparation of glass-ceramic materials, which represent an effective option to manage hazardous waste. In this study, both preliminary spinel and secondary spinel were precipitated in the precursor glass. The formation of these spinel was meticulously assessed by a combination of X-ray diffractometry and scanning electron microscopy. The structure of the microenvironment in the precursor glass was characterized by Raman spectrums. These advanced techniques highlight the potential for one-step crystallization of the glass. The investigation, which focused on one-step crystallization, demonstrated the growth of pyroxene on spinel accompanying a migration of chromium. The results also show the microstructure of the obtained glass-ceramic was very dependent on the heat-treat temperature. This study not only unambiguously reveals the precipitation mechanisms of spinel but also provides more documentation for one-step crystallization in the glass-ceramics field. 相似文献
49.
《Intermetallics》2015
Fine-grained fully-lamellar (FL) microstructure is desired for TiAl components to serve as compressor/turbine blades and turbocharger turbine wheels. This study deals with the process and phase transformation to produce FL microstructure for Mo stabilized beta-gamma TiAl alloys without single α-phase field. Unlike the α + γ two-phased TiAl or beta-gamma TiAl with single α-phase field, the wrought multi-phase TiAl–4/6Nb–2Mo–B/Y alloys exhibit special annealing process to obtain FL microstructure. Short-term annealing at temperatures slightly above β-transus is recommended to produce the desired FL microstructure. The related mechanism is to guarantee the sufficient diffusion homogenization of β stabilizers during single β-phase annealing, and further avoid α decomposition by α → γ + β when cooling through α + β + γ phase field. The colony boundary β phase contributes to fine-grained nearly FL microstructure, by retarding the coarsening of the α phase grains. 相似文献
50.