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71.
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
72.
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
73.
74.
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
75.
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77.
汽车用塑料注塑新技术的进展 总被引:1,自引:0,他引:1
综述了汽车用塑料的注射成型技术的进展,简单介绍了气体辅助注射成型(GAIM)、水辅助注射成型、熔芯注射成型、反应注射/增强反应注射(RIM/RRIM)成型等技术的特点、生产的汽车部件及最新动向。 相似文献
78.
β-SiC powder containing 6 wt% A12 O3 and 4 wt% Y2 O3 as sintering additives was pressureless sintered at 2000°C for 1 h (AYE-SiC) and 3 h (AYP-SiC). AYE-SiC consisted of an equiaxed grain structure and AYP-SiC exhibited a micro-structure with platelike grains as a result of grain growth related to β→α phase transformation during sintering, R -curve behavior and flaw tolerance for these silicon carbides were evaluated by the indentation-strength technique. For comparison, the R -curve behavior of conventional sintered, boron- and carbon-doped SiC (SS-SiC) was evaluated. AYE-SiC and AYP-SiC exhibited rising R -curve behavior with toughening exponents of m = 0.042 and m = 0.135, respectively. AYP-SiC exhibited better flaw tolerance and more sharply rising R -curve behavior than AYE-SiC. The more sharply rising R -curve behavior and the better flaw tolerance of AYP-SiC were attributed mainly to grain bridging of crack faces by platelike grains. Because of the high degree of transgranular fracture, SS-SiC exhibited a flat R -curve despite a microstructural feature with platelike grains. 相似文献
79.
Yongwon Seo Sung-Ho Jo Ho-Jung Ryu Hee Dal Bae Chong Kul Ryu Chang-Keun Yi 《Korean Journal of Chemical Engineering》2007,24(3):457-460
A bubbling fluidized bed reactor was used to study CO2 capture from flue gas by using a potassium-based solid sorbent, sorbKX35 which was manufactured by the Korea Electric Power
Research Institute. A dry sorbent, sorbKX35, consists of K2CO3 for absorption and supporters for mechanical strength. To increase initial CO2 removal, some amount of H2O was absorbed in the sorbent before injecting simulated flue gas. It was possible to achieve 100% CO2 removal for more than 10 minutes at 60°C and a residence time of 2 s with H2O pretreatment. When H2O pretreatment time was long enough to convert K2CO3 of sorbKX35 into K2CO3 · 1.5H2O, CO2 removal was excellent. The results obtained in this study can be used as basic data for designing and operating a large scale
CO2 capture process with two fluidized bed reactors.
This work was presented at the 6
th
Korea-China Workshop on Clean Energy Technology held at Busan, Korea, July 4–7, 2006. 相似文献
80.
Jae Ryong Kim Do-Hyeong Kim Chong Hee Kim 《Journal of the American Ceramic Society》1990,73(8):2567-2569
The effects of ZrO2 and Y2 O3 on the densification of hotpressed Si3 N4 -Zr(Y)O2 composites have been studied. High density could not be obtained by the addition of pure or 3-mol%-Y2 O3 -doped ZrO2 in this composite; however, nearly full density (>97%) could be obtained in Si3 N4 using 6- and 8-mol%-Y2 O3 -doped ZrO2 . It is concluded that Y2 O3 diffusing out from the added Zr(Y)O2 promoted the densification and that ZrO2 also had some role in the formation of an oxynitride glass. 相似文献