The aim of study To purify GPCR from a local strain of S. cerevisiae using Ion exchange and gel filtration chromatography techniques , by packing materials for columns which will be chosen of low cost comparing to the already used in published researches, which depend on the costly affinity chromatography and other expensive methods of purification. Local strain of S. cerevisiae chosen for extraction and purification of G-protein coupled receptor (GPCR) .The strains were obtained from biology department in Al- Mosul University, Iraq. The isolated colony was activated on Yeast Extract Pepton Dextrose Broth (YEPDB) and incubated at 30 C˚ for 24 h .Loop fully of the yeast culture was transferred to (10ml) of yeast extract peptone glucose agar (YEPGA) slant , then incubated at 30C˚for 24h , after that it was stored at 4C˚ ,the yeast cultures were reactivated and persevered after each two weeks period. S.cerevisiae was identified by morphological, microscopic characterization and biochemical test . The GPCR that extract from whole cell of S.cerevisiae was purified by ion exchange chromatography using DEAE-Sepharose ,the bound proteins (negatively charged) were then eluted using gradient concentration of NaCl ranged between( 0.1 -0.5M). Gel filtration chromatography using Sepharose 6B was applied as a second step of purification. The optical density for each fraction was measured at 280 nm by UV-VS spectrophotometer then the GPCR concentration was determined by using ELISA Kit . The fractions which gave the highest absorbance and concentration of GPCR were collected .The molecular weight of GPCR was determined by gel filtration chromatography using blue dextrin solution. Standard curve was plotted between log of molecular weight for standard protein and the ratio of Ve/Vo of GPCR . The purity of the GPCR that extracted and purified from whole cell of S, cerevisiae were carried out by using SDS-PAGE electrophoresis . In ion exchange chromatography the fraction were collected with 5 ml tube at a flow rate 0.5 ml/ min and eluted with gradient (0.1-0.5M) of sodium chloride solution. Two proteins peaks appeared after eluted by the gradient concentration of sodium chloride, while no protein peaks appeared in the washing fractions. The GPCR concentration was measured in the fractions of these two protein peaks, data indicated that GPCR located in the first protein peak (eluted at 0.1M of NaCl) at fraction numbers between 3 and 9, the maximum concentration of GPCR was 9.281 with specific activity 71.58(ng/mg)protein , 3.125 purification folds and72.9(%) yield while the second peaks (eluted at 0.4 M of NaCl) don't give any concentration for GPCR, thus its neglected. Gel filtration chromatography was used as second step of purification which applied by using sepharose 6B. Results show single active protein peaks appeared that identical with the peak of GPCR at fractions numbers(29-35). The maximum concentration of GPCR was 9.082 (ng/ml)was observed in these fractions. The specific activity for these fractions was 151.37 (ng/mg) protein with 6.608 purification folds and 39.64 (%) yield. The present study a chive a relatively high purification of GPCR from whole cell of a local strain S. cerevisiae with fold purification 6.608 and a yield of 39.64 % and molecular weight about~33KD.
Acute lymphoblastic leukemia (ALL) is a cancer of the blood and bone marrow (spongy tissue in the center of bone). In ALL, too many bone marrow stem cells develop into a type of white blood cell called lymphocytes. These abnormal lymphocytes are not able to fight infection very well. The aim of this study was to investigate possible links between E3 SUMO-Protein Ligase NSE2 [NSMCE2] and increase DNA damage in the childhood patients with Acute lymphoblastic leukemia (ALL). Laboratory investigations including hemoglobin(Hb) ,white blood cell (WBC) , serum total protein , albumin ,globulin , in addition to serum total antioxidant activity (TAA) , Advanced oxidation protein products(AOPP) and E3 SUMO-Protein Ligase NSE2[NSMCE2]. Blood samples
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The CIGS/CdS p-n junction thin films were fabricated and deposited at room temperature with rate of deposition 5, and 6 nm secG1 , on ITO glass substrates with 1mm thickness by thermal evaporation technique at high vacuum pressure 2×10G5 mbar, with area of 1 cm2 and Aluminum electrode as back contact. The thickness of absorber layer (CIGS) was 1 µm while the thickness of the window layer CdS film was 300 nm. The X-ray Diffraction results have shown that all thin films were polycrystalline with orientation of 112 and 211 for CIGS thin films and 111 for CdS films. The direct energy gaps for CIGS and CdS thin films were 1.85 and 2.4 eV, respectively. Atomic Force Microscopy measurement proves that both films CIGS and CdS films have nanostru
... Show MoreIn order to have an idea about what happens in Iraqi food establishments in relation to implement quality management system ISO 9001, this study was performed to show the actual situation of Iraqi food establishments concerning quality management system (ISO 9001:2015), reasons of implementing, factors that hinder implementing and problems faced high administration for getting establishments certification ISO 9001:2015. The study demonstrated from the questionnaire some difficulties to implement ISO 9001 for both of establishments that implemented the quality system or which in implementing of this international standard. The most important problems during implementing were business culture and costs and the most important proble
... Show MoreThe appearance of Mixed Mode Oscillations (MMOs) and chaotic spiking in a Light Emitting Diode (LED) with optoelectronic feedback theoretically and experimentally have been reported. The transition between periodic and chaotic mixed-mode states has been investigated by varying feedback strength. In incoherent semiconductor chaotically spiking attractors with optoelectronic feedback have been observed to be the result of canard phenomena in three-dimensional phase space (incomplete homoclinic scenarios).
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In this work, the fusion cross section , fusion barrier distribution and the probability of fusion have been investigated by coupled channel method for the systems 46Ti+64Ni, 40Ca+194Pt and 40Ar+148Sm with semi-classical and quantum mechanical approach using SCF and CCFULL Fortran codes respectively. The results for these calculations are compared with available experimental data. The results show that the quantum calculations agree better with experimental data, especially bellow the Coulomb barrier, for the studied systems while above this barrier, the two codes reproduce the data.
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A model using the artificial neural networks and genetic algorithm technique is developed for obtaining optimum dimensions of the foundation length and protections of small hydraulic structures. The procedure involves optimizing an objective function comprising a weighted summation of the state variables. The decision variables considered in the optimization are the upstream and downstream cutoffs lengths and their angles of inclination, the foundation length, and the length of the downstream soil protection. These were obtained for a given maximum difference in head, depth of impervious layer and degree of anisotropy. The optimization carried out is subjected to constraints that ensure a safe structure aga
... Show MoreThis paper is concerned with the blow-up solutions of a system of two reaction-diffusion equations coupled in both equations and boundary conditions. In order to understand how the reaction terms and the boundary terms affect the blow-up properties, the lower and upper blow-up rate estimates are derived. Moreover, the blow-up set under some restricted assumptions is studied.
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Heat shock protein 70 (HSP70) is a crucial protein with vital biological tasks in cell continuation of life. The variation of HSP70 activation occurs as a consequence of stress that includes temperature states, toxicity, poisoning with heavy metals, and tumor-related conditions. One of the master jobs of the HSP family is the suppression of caspase-mediated apoptosis signals. A high level of the expression of HSP70 is accountable for tumorigenesis and resistance against chemotherapeutic drugs. For this reason, the detection of HSP70 may help to diagnose cancerous diseases. From the other side, targeting this chaperone might help in treatment by maintaining late caspase-dependent events. This study was conducted to detect the presenc
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