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.
KE Sharquie, AA Noaimi, Journal of the Saudi Society of Dermatology & Dermatologic Surgery, 2012 - Cited by 36
Structural and optical properties were studied as a function of Nano membrane after prepared, for tests. Nano membrane was deposited by the spray coating method on substrates (glass) of thickness 100 mm. The X-ray diffraction spectra of (CNTs, WO3) were studied. AFM tests are good information about the roughness, It had been designed electrolysis cell and fuel cell. Studies have been performed on electrochemical parameters.
This study has been carried out in the animal field of the college of agricultural engineering sciences, university of Baghdad, for the period from 12/15/2021 to 1/26 /2022 for 42 d, to investigate the effect of adding different levels of ellagic acid to the diet of broilers, on some physiological characteristics & oxidation indicators in meat compared to vitamin C in meat, 225 Ross 308 chicks were used, divided randomly to five treatments such us: T1: control group without additives to diet, & the other T2, T3, T4 was added ellagic acid (
... Show MoreBackground: The study was designed to evaluate the effect of local application of exogenous VEGF/collagen I separately and as a combination in socket healing. Sixty male Albino Wistar rats were subjected for a surgical tooth extraction of upper 1st molar of both sides (right side was considered as experimental site, while left be the control one, treated with 1µL of normal saline). The rats were scarified at 3, 7, 14, 28 days post extraction. Socket healing was histologically examined with immunohistochemistochemical localization of ALP&FGF2. Materials and Method: Sixty male Albino Wistar rats were subjected for a surgical tooth extraction of upper 1stmolar of both sides (right side was considered as experimental site, while left be the
... Show MoreThe main aim of this study is to investigate the ability of four local entomopathogenic isolates Beauveria bassiana (Bals.) and Metarhizium anisopliae (Met.) to control the mosquito larvae in the lab. The results revealed that the isolate (MARD48) B .bassiana reduced the survival rate of the mosquito larvae to (80%) followed by the isolate M. anisopliae (MARD10) to (90%) in the first two days of treatment, and 60 and 66% respectively in the third day. The results also showed that the isolate B. bassiana (MARD48) killed 50% of the population (LC50) with the concentration 1×104 conidia/ml compared to 1×107 conidia/ml for the isolates B. bassiana (MARD14) and M. anisopliae (MARD10), and 1×108 conidia/ml for the isolate B .bassiana (MARD76).
... Show MoreIn this research, we studied the effect of concentration carriers on the efficiency of the N749-TiO2 heterogeneous solar cell based on quantum electron transfer theory using a donor-acceptor scenario. The photoelectric properties of the N749-TiO2 interfaces in dye sensitized solar cells DSSCs are calculated using the J-V curves. For the (CH3)3COH solvent, the N749-TiO2 heterogeneous solar cell shows that the concentration carrier together with the strength coupling are the main factors affecting the current density, fill factor and efficiency. The current density and current increase as the concentration increases and the
Background: Bone is essentially a highly vascular, living, constantly changing mineralized connective tissue. It is remarkable for its hardness, resilience and regenerative capacity, as well as its characteristic growth mechanisms. This study aimed to: 1. To evaluate the effect of bone morphogenetic protein7 (BMP7) on bone healing in artificially created intrabony defect in rabbits upper diastema, histologically. 2. To study the immunohistochemical expression of TGF-β3 and IGF-1R as bone formation markers in experimental and control groups during bone healing. Material and method: Forty male rabbits, was used in this study, 8 rabbits for each healing interval (3 days, 1,2 ,4 and 6 weeks). In each rabbit two bone holes were created on th
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