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1.
Expression,purification and spectra characterization of neuroglobin   总被引:1,自引:0,他引:1  
The expression, purification and spectra characterization of recombinant human neuroglobin (NGB) are reported. The pET3a plasmid with the gene of NGB was transformed to E. coli BL21 (DE3) plys cells and expressed in TB culture medium. The results indicated that the expression amount of NGB is about 10 percent of the total protein in cells. The NGB protein was purified by ammonium sulfate precipitation, DEAE-Sepharose anion exchange column, Hiload 16/60 superdex 75 size exclusion chromatography and a Hiprep 16/10 Q FF anion exchange column, and a red soluble protein was obtained which showed a single band in electrophoresis. Electrospray ionization mass spectrometry (ESI-MS) showed that its molecular weight is 16930.0 Da. UV-spectra indicated that the reduced NGB has a strong absorption peak at 425 nm, and two weak peaks at 531 and 559 nm, which can be assigned to γ, β and α bands of porphyrin, respectively, and the oxidized NGB has a strong absorption peak at 413 nm which corresponds to the transition of π electrons in the porphyrin ring. The fluorescence maximal excitation wavelength is at 281 nm and its maximal emission wavelength is at 338 nm. CD spectra indicated that its secondary structure is a typical α helix, and has a positive peak at 410 nm induced by heme, The NGB protein is stable when the pH is higher than 4.  相似文献   

2.
The variable domain of heavy chain (VH) and light chain (VL) genes of anti-CD20 monoclonal antibody HI47 were cloned from anti-CD20 ScFv expression vector pCANBTEcd20 by PCR and ligated into vector pYZF to construct chimeric anti-CD20 Fab’ fragment expression vector pYZFcd20. Chimeric anti-CD20 Fab’ fragment was expressed inE. coli 16C9 and purified by protein G affinity chromatography. Competitive inhibition assay showed that anti-CD20 Fab’ fragment inhibited binding of HI47 to CD20 on the surface of Daudi cells. Results from MTT assay indicated that chimeric anti-CD20 Fab’ fragment inhibited the proliferation of Daudi cells, IC50 = 69 μg/mL. Affinity of chimeric anti-CD20 Fab’ fragment was determined, Ka was about 8.9×108 (mol/L)−1.  相似文献   

3.
HARMFUL ALGAL BLOOM ACTS UPON IMPORTANT ENVIRON-MENTAL PROBLEMS IN COASTAL WATERS.A UNIFIED AND IM-PLEMENTING METHOD FOR SURVEY AND MONITORING ALGALBLOOMS HAS BEEN DEVELOPED SINCE LAST DECADE.HOWEVER THERE IS A DATA GAP OF ABOUT TENS YEARS IN THE PERIOD W…  相似文献   

4.
5.
Activities and properties of calcineurin catalytic domain   总被引:2,自引:0,他引:2  
Calcineurin (CN) is the only protein phosphatase known to be under the control of calcium (Ca2+) and calmodulin (CaM). The enzyme consists of two subunits, the catalytic A subunit of 61 ku (CNA) and a regulatory B subunit of 19 ku (CNB). In this study, we used PCR amplication to construct a truncation consisting of only the CNA catalytic domain. The truncation was induced by IPTG and expressed inE. coli. PNPP was used as a substrate to study the phosphatase activity of the CNA catalytic domain. The findings show that its activity is 20 times greater than CNA in the presence of CNB and CaM. The optimum reaction temperature for the CNA catalytic domain protein is 40°C, and the optimum reaction pH value is 8.0. Mn2+ is still an effective activator for the CNA catalytic domain, but its activity is not controlled by Ca2+. In the presence of 6 mmol/L Mg2+, adding either Ca2+ or EGTA did not change the activity of the CNA catalytic domain.  相似文献   

6.
Pulsed laser photolysis/laser-induced fluorescence (LP-LIF) is utilized to measure rate constants for C2(a3Πu) reactions with NO, N2O, O2, H2 and NH3. Multiphoton dissociation of C2Cl4 at 266 nm is employed for the generation of C2(a3Πu) radicals. The C2(a3Πu) concentration is monitored by the fluorescence of the (0, 0) band of the (d3Πg&#8596;a3Πu) transition at 516.5 nm. C2(a3Πu) removal rate constants for the reactions are determined as kNO = (5.46 ± 0.10) × 10-11 cm3 molecule-1 s-1 , kN2O = (1.63 ± 0.20) × 10-13 cm3 molecule-1 s-1 , kN2O = (1.58 ± 0.16) × 10-11 cm3 molecule-1 s-1, kO2 = (5.92 ± 1.00) × 10-14 cm3 molecule-1 s-1, kH2< 1.0× 10-14 cm3 molecule-1 s-1. Based on the data analysis and theoretical calculation, we suggest that the C2(a3Πu) reactions with H2 and NH3 proceed via the hydrogen abstraction mechanism, barriers exist at the entrance channel of the reactions of C2(a3Πu) with H2 and NH3.  相似文献   

7.
The photoluminescence quenching behaviors of ^5D3-^7Fj and ^5D4-^7Fj (J = 0—6) transitions of Tb^3+ in YBO3:Tb under 130—290 nm excitation were systematically investigated. The results revealed that the quenching concentrations of both ^5D3-^7Fj and ^5D4-^7Fj transitions of Tb^3+ in YBO3:Tb were mainly dependent on excitation wavelength. Particularly, the quenching concentrations of ^5D4-^7Fj transitions of Tb^3+ under 130—290 nm excitation were correlated with excitation bands of YBO3:Tb. The quenching concentrations of ^5D3-^7Fj transitions remained at low concentration (2%) under 186—290 nm excitation and then increased gradually with energy of incoming excitation photon when excited at 130—186 nm. This dependence should be involved in their excitation mechanisms and quenching pathway in particular excitation region.[第一段]  相似文献   

8.
二维(2D)过渡金属碳/氮化物(MXene)材料是当前最受关注的二维材料之一,其中二维碳化钛(Ti3C2Tx MXene)材料的研究最为广泛。该材料目前主要通过刻蚀三元碳化物或氮化物(MAX相)后进一步插层得到,因此MAX相材料的纯度和制备工艺条件直接决定了Ti3C2Tx MXene材料的物化性质。主要完成了不同Ti3AlC2 MAX相材料的筛选,选择氢氟酸(HF)刻蚀,并优化了不同的插层方法,制备了一系列Ti3C2Tx MXene材料。通过X射线衍射(XRD)、场发射扫描电镜(FESEM)、透射电子显微镜(TEM)、原子力显微镜(AFM)和X射线光电子能谱(XPS)等表征,确定使用原位锂离子(Li+)插层法可有效获得单层Ti3C2Tx MXene材料。制备的单层Ti3C2Tx MXene材料的表面平整,片径约为150 nm,厚度约为2 nm。同时,创新性地采用涡旋震荡辅助材料分层,极大地缩短了超声时间,提高了单层Ti3C2Tx MXene材料的产率(可达70%),并且可以避免材料氧化,为Ti3C2Tx MXene材料未来应用提供了新方法。  相似文献   

9.
Nanometer sized Au/TiO2, particles were synthesized by irradiation of a HAuCl4 solution containing colloidal TiO2 with light of wavelength (λ) > 330 nm. The absorption maximum attributed to the surface plasmon band of gold was observed at 540 nm, a red shift of about 20 nm from the position in aqueous solution. The Au clusters are situated on the surface of TiO2 in terms of microcrystallite, which was confirmed by HRTEM, EDS and XRD. The electronic interfaction between the metal and the suport was discussed.  相似文献   

10.
Two ternary complexes Cu2A4[OP(OCH3)3]2 (A represents CH2== CH—COO- and CH2== C(CH3)—COO-) have been synthesized, and elemental analyses, IR, ESR, electronic reflectance spectra and magnetic studies were carried out. The single crystal X-ray diffraction shows that Cu2[CH2== C(CH3)—COO]4[OP(OCH3)3]2 is triclinic, with space group P1, a = 1.05128(13), b = 1.7559(5), c = 1.94479(3) nm, α = 91.263(14)°, β = 102.559(6)°, γ = 106.339(13)°, Z = 4 and R = 0.0668. Two copper(Ⅱ) atoms are bridged by four a -methacrylate groups, and each copper(Ⅱ) atom is coordinated with a trimethyl phosphate molecule in the axial position, forming a distorted square pyramidal configuration. The symmetric center is between the two copper(Ⅱ) atoms, and the Cu-Cu bond distance is 0.26098(6) nm. The Cu-Cu distance and magnetic studies suggest that there exist antiferromagnetic interactions between the two copper(Ⅱ) atoms.  相似文献   

11.
运用水样叶绿素荧光仪(WATER-PAM)研究Cu2+胁迫下,湛江叉鞭金藻(Dicrateria zhanjiangensis)和球等鞭金藻3011(Isochrysis galbana MACC/H59)叶绿素荧光特性的变化,同时监测微藻的生长情况.结果就叶绿素荧光参数而言,湛江叉鞭金藻和球等鞭金藻3011的叶绿素荧光参数Fv/Fm,Fv/F0,Yield和ETR均随着Cu2+浓度的增大而明显降低;就生长情况而言,随着Cu2+浓度的增大,两种海洋微藻细胞密度的增长明显变缓.在24 h时Cu2+对湛江叉鞭金藻的毒性大于球等鞭金藻3011,48~96 h时Cu2+对球等鞭金藻3011的毒性大于湛江叉鞭金藻,Cu2+的毒性随着胁迫时间的延长而逐步增大.  相似文献   

12.
We explore nitric oxide (NO) effect on K^+in, channels in Arabidopsis guard cells. We observed NO inhibited K^+in, currents when Ca^2+ chelator EGTA (Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N;tetraacetic acid) was not added in the pipette solution; K^+in currents were not sensitive to NO when cytosolic Ca^2+ was chelated by EGTA. NO inhibited the Arabidopsis stomatal opening, but when EGTA was added in the bath solution, inhibition effect of NO on stomatal opening vanished. Thus, it implies that NO elevates cytosolic Ca^2+ by activating plasma membrane Ca^2+ channels firstly, then inactivates K^+in, chartnels, resulting in stomatal opening suppressed subsequently.  相似文献   

13.
 采用HPLC法测定五香血藤中五内酯E的含量.色谱柱:sphenrisorb C18(4.0 mm×250 mm,5μm),流动相:A水,B乙腈,按体积分数梯度洗脱70%~90%,0~25 min,波长210 nm.线性范围为1.00~5.00μg·mL-1,相关系数R2=0.999 9,平均回收率为97.73%,相对标准偏(RSD)为2.33%.方法具有简単、灵敏、准确等特点.使用该方法测定了贵州8个不同产地的五香血藤的含量,结果满意.  相似文献   

14.
After removal of cytoplasmic sector F1 from submitochondrial particles of F0F1-ATP synthase complex with guanidine hydrochloride, the transmembrane sector F0 was specifically extracted from the stripped membranes in the presence of detergent CHAPS and partially purified. Two-dimensional crystals were produced by the reconstitution of the partially purified F0 into asolectin and microdialysis. The obtained crystals are able to diffract to 2 nm. The projection map of the negatively stained crystal shows that the crystal has p4212 symmetry, lattice constant, a=b=14.4 nm. A unit cell contains four F0 molecules.  相似文献   

15.
Two binuclear copper(Ⅱ) complexes, [Cu2(saloph)2 (μ-O)2]-2(DMF)[H2saloph=N,N′-o-phenylenebis(salicylideneaminato)] (a) and [Cu2(salen)2(μ-O)2] [H2salen=N,N′-bis(salicylic-deneaminato) ethylene] (b)were synthesized and characterized by X-ray crystallography. Both of them have distorted rectangular pyramidal geometry around Cu(Ⅱ). The complete series of complexes show bridging phenoxo groups between the copper centers, together with hydroxo-bridges in these complexes. The complexes have also been characterized by elemental analysis, IR, TG-DTA, and electrochemical results.  相似文献   

16.
The relationship between the stomatal density of five woody plants endemic to China, i. e.Eucommia ulmoides, Quercus liaotungensis, Q. glandulifera var.brevipetiolata, Cyclocarya paliurus andFicus heteromorpha, and the atmospheric CO2 concentrations was studied by observations on leaves of the herbarium-stored specimms(1920s–1990s). The results showed that the stomatal density inEucommia ulmoides, Quercus liaotungensis andQ. glandulifera var.brevipetiolata decreased significantly in response to the elevated atmospheric CO2 concentrations, while inCyclocarya paliurus it decreased slightly and inFicus heteromorpha there were no responses.  相似文献   

17.
Mg base BMG alloys have attracted a great deal of attention for its low density and relative low cost. To date, BMG alloys with millimeter scale have been fab- ricated by using conventional copper cast and die cast method in Mg-Ni-Ce[1], Mg-Ni-La[2], Mg-C…  相似文献   

18.
A series of sandwich-type tungstoarsenates heteropoly compounds with As/W ratio of 4/30, Na16[As4W30M4(H2O)2O112]·XH2O (M = Zn,Cu,Co,Ni,Mn and Cd), have been synthesized for the first time and structurally characterized by elemental analysis, IR and 183W NMR spectra. The crystal structure of Na16[As4W30Cu4(H2O)2O112]·63H2O was determined to be a triclinic system, of P1 symmetry, a = 1.2721(3) nm, b = 2.451 6(5) nm, c = 2.6450(5) nm, α= 89.90(3)°,β= 77.32(3)°, γ= 89.96(3)°, 2=2. Using tetrahepty lammonium bromide as a phase transfer reagent, [As4W30Cu4(H2O)2O112]16- was transferred from aqueous solution to organic phase (benzene), and the heteropolyanion lost the coordination water molecules to form the coordination-unsaturated ion. After lactic acid was added to the benzene solution, the coordination-saturation was recovered. By esterification reaction between lactic acid and cholesterin, the latter was attached to the heteropolyanion indirectly. Therefore, a new type of lyotropic liquid crystal was obtained, which was characterized by a polarimicroscope, DSC and variable temperature wide-angle X-ray diffraction.  相似文献   

19.
The structural parameters of nine Indian coals were determined by X-ray diffraction (XRD) and Raman spectroscopy. The study revealed that the coals contain crystalline carbon of turbostratic structure with amorphous carbon. The stacking height (Lc) and interlayer spacing (d002) of the crystallite structure of the coals ranged from 1.986 to 2.373 nm and from 0.334 to 0.340 nm, respectively. The degree of graphitization was calculated to range from 42% to 99%, thereby confirming the ordering of the carbon layers with the increase in coal rank. An exponential correlation was observed among the aromaticity (fa), the lateral size (La), and the rank (I20/I26), suggesting that the coal crystallites are nanocrystalline in nature. A very strong correlation was observed between the structural parameters (fa, d002, Lc, the H/C ratio, and I20/I26), the volatile matter content, and the elemental carbon content, indicating the structures of coals are controlled by the degree of contact metamorphism. The Raman spectra exhibited two prominent bands: the graphitic band (G) and the first-order characteristic defect band (D). The deconvolution resulted in five peaks: G, D1, D2, D3, and D4. The intense D1 band, which appeared at ~1350 cm?1, corresponds to a lattice vibration mode with A1g symmetry. The D2 mode, which appeared at ~1610 cm?1, arises from the structural disorder as a shoulder on the G band.  相似文献   

20.
Fe+ ion beams with the energy of 110 keV were implanted into films of L(+)-cysteine (HSCH2CH(NH2)COOH). One of the single crystals grown in hydrochloric acid solution with the implanted samples through slow evaporation was structurally characterized by the X-ray crystallography. The crystal is monoclinic, space group C2, with a = 1.8534(4) nm, b = 0.5234(1) nm, c = 0.7212(1) nm, β= 103.72°, V = 0.67965(3) nm3, Z = 4, F(000) = 144.0, D{clac} = 1.763 g · cm−3, μ(MoK a = 1.06 mm−1, T = 293(2) K. R = 0.0379, wR = 0.0835 for 660 observed reflections (I > 2σ(I)). The structural formula of the crystal compound is (CH2CH(NH2)NO2)ClFe (M r = 180.38 u). Products of heavy ion beam irradiation were purified and it was directly confirmed that the implanted Fe+ ions had been deposited in the novel molecules. The same doses of Fe+ ion beams of the same energy were implanted into films of L(+)-cysteine hydrochloride monohydrate. FTIR spectroscopy of the implanted samples proved that some of the original molecules were seriously damaged and significant modifications were induced.  相似文献   

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