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.
The research aims to investigate the existence of a direct causal relationship between selected agricultural variables: agricultural output (as a representative of growth in the agricultural sector), agricultural terms of trade as a completely new variable in agricultural studies in recent years, agricultural labour which is an important part in the total workforce for Iraq, and finally, agricultural investment because of its importance and vital role in the production process, creating job opportunities, and then raising the level of employment, then it's role to achieving agricultural growth and development. For this purpose, the researchers used the Toda-Yamamoto causality methodology for a time series covering from 1990 to 2019. The res
... Show MoreMn2O3 was coated onto reduced titania nanotubes by reverse pulse electrodeposition, showing smooth and homogenous deposits without covering the opening of the nanotubes.
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The impact of a Schiff base namely 2-((thiophen-2-ylmethylene)amino)benzenethiol to corrode mild steel in 1 M HCl resolved was evaluated using different weight loss technique and scanning electron microscopy (SEM).different weight measurements to expand that the 2-((thiophen-2-ylmethylene) amino) benzenethiol inhibits the corrosion of mild steel through adsorbing of top for mild steel and block the active locality. The inhibitive impacts of 2-((thiophen-2-ylmethylene)amino)benzenethiol increase with increasing concentration and decrease with increasing temperature. SEM to checking revealed that the alloy surface was quite unaffected and formed protective film on its surface. The investigated
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Irrigation has significant role in endodontic treatment, many types of antimicrobial irrigation solutions have been used, but due to the ineffectiveness, safety concerns and side effects of this irrigation, the herbal alternatives for endodontic irrigants might be beneficial. Objectives This study compared the in vitro effectiveness of tea tree oil and clove oil as possible irrigants in endodontics against Enterococcus faecalis in comparison with 3% Sodium hypochlorite. Materials and Methods E. faecalis was isolated from patients in need for endodontic treatment; VITEK was employed for E. faecalis isolate conformation. Muller Hinton agar was prepared with 100μl of freshly prepared suspension of E.faecalis. Wells of 6mm diameter and 4mm dep
... Show MoreOne of the most common public liver diseases over the world is fatty liver which contain alcoholic and non-alcoholic fatty liver. One-fourth among general population are impact Non-Alcoholic Fatty Liver Disease (NAFLD) in the worldwide.Retinol binding protein 4 (RBP4) is known as an adipokine, mainly synthesized and secreted from the liver and form adipose tissues. RBP4 acts as a transporter and specifically bound to retinol from liver to others tissues. Visfatin is an adipocytokine and mainly produced from visceral fat tissue, skeletal muscles as well as liver. Vitamin A absorbed, transported as retinyl esters to the liver then hydrolyzed to the retinol form and storage in hepatic stellate cells (HSCs) after reesterified with rigly
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This study aimed at revealing the degree of availability of standards of word problems in mathematics books for the first three grades of the basic stage in Palestine. For this purpose, the researcher prepared an analysis tool and a list of criteria consisting of two areas: linguistic formulation and mathematical content. Every area had seven items. The results of the study showed that the third-grade mathematics book has the highest degree of availability of the standards with 85.75%, and then came the second-grade mathematics book with 83.12%. Finally, the first-grade mathematics book came with 80.13%. In the light of the previous results, the researcher recommended to develop the language of word problems, to take into account their i
... Show MoreSince the COVID-19 pandemic alarm was made by the severe acute respiratory syndrome (SARS)-coronavirus (CoV) 2, several institutions and agencies have pursued to clarify the viral virulence and infectivity. The fast propagation of this virus leads to an unprecedented rise in the number of cases worldwide. COVID-19 virus is exceptionally contagious that spreads through droplets, respiratory secretions, and direct contact. The enveloped, single-stranded RNA virus has a specific envelop region called (S) region encoding (S protein) that specifically binds to the host cell receptor. Viral infection requires receptors' participation on the host cell membrane's surface, a key- step for the viral invasion of susceptible cells.
Rec
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Abstract: New copper(II) complexes with mixed ligand benziloxime (BOxH) and furfural-dehydeazine (FA) using classical (with and without solvent) and microwave heating methods have been prepared. The resulting complexes have been characterized using physico-chemical techniques. The study suggested that the ligands formed neutral complexes had general formulas [Cu(FA)(BOXH)(Ac)2] and [Cu(FA)(BOX)(OH)] in neutral (or acidic) and basic medium, respectively. Accordingly, hexa-coordinated mono-nuclear complexes have been investigated by this study and having distorted octahedral geometry. The effect of laser have been studied on solid ligands and solid complexes, no effect have been observed on most compounds through the results of melting poin
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Background: The present in-vitro study was undertaken to evaluate and compare fracture resistance of weakened endodontically treated premolars with class II MOD cavities restored with different bulk fill composite restorations (EverX posterior, Alert, Tetric EvoCeram Bulk Fill, and SDR). The type and mode of fracture were also assessed for all the experimental groups. Materials and Method: Forty-eight human adult maxillary premolar teeth were selected for this study. Standardized extensive class II MOD cavities with endodontic treatment were prepared for all teeth, except those that were saved as intact control. The teeth were divided into six groups of eight teeth each (n=8): (Group 1) intact control group, (Group 2) unrestored teeth with
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