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.
This paper including a gravitational lens time delays study for a general family of lensing potentials, the popular singular isothermal elliptical potential (SIEP), and singular isothermal elliptical density distribution (SIED) but allows general angular structure. At first section there is an introduction for the selected observations from the gravitationally lensed systems. Then section two shows that the time delays for singular isothermal elliptical potential (SIEP) and singular isothermal elliptical density distributions (SIED) have a remarkably simple and elegant form, and that the result for Hubble constant estimations actually holds for a general family of potentials by combining the analytic results with data for the time dela
... Show MoreThe present work utilizes polyacrylic acid beads (PAA) to remove Alizarin yellow R (AYR)] and Alizarin Red S (ARS) from its solution. The isotherms of adsorption were investigated and the factors that impact them, such as temperature, ionic strength effect, shaking effect, and wet PAA. The isotherms of adsorption of (ARS) were found obeys the Freundlich equation. The isotherms of adsorption of (AYR) were found obeys the Langmuir equation. At various temperatures, the adsorption process on (PAA) was investigated. According to our data, there is a positive correlation between the (ARS and AYR) adsorption on the PAA and temperature (Endothermic process). The computation of the thermodynamic functions (ΔH, ΔG, and ΔS) is based on the foregoi
... Show MoreFace recognition, emotion recognition represent the important bases for the human machine interaction. To recognize the person’s emotion and face, different algorithms are developed and tested. In this paper, an enhancement face and emotion recognition algorithm is implemented based on deep learning neural networks. Universal database and personal image had been used to test the proposed algorithm. Python language programming had been used to implement the proposed algorithm.
Lithology identification plays a crucial role in reservoir characteristics, as it directly influences petrophysical evaluations and informs decisions on permeable zone detection, hydrocarbon reserve estimation, and production optimization. This paper aims to identify lithology and minerals composition within the Mishrif Formation of the Ratawi Oilfield using well log data from five open hole logs of wells RT-2, RT-4, RT-5, RT-6, and RT-42. At this step, the logging lithology identification tasks often involve constructing a lithology identification model based on the assumption that the log data are interconnected. Lithology and minerals were identified using three empirical methods: Neutron-Density cross plots for lithology id
... Show MoreOut of 150 different specimens, 67 S. aureus isolate were isolated. However, 16sRNA gene was located only in 60 isolates. Moreover, mecA gene was located in 48 isolates; thereby MRSA covered 80% of all S. aureus isolates. Of considerable interest, pvl gene was detected in only six isolates (10%). Hence, the present work emphasizes the notion suggested that pvl is not an indicative of CA-MRSA.
Derivatives of Schiff-bases possess a great importance in pharmaceutical chemistry. They can be used for synthesizing different types of bioactive compounds. In this paper, derivatives of new Schiff bases have been synthesized from several serial steps. The acid (I) was synthesized from the reaction of dichloroethanoic acid with 2 moles of p-aminoacetanilide. New acid (I) converted to its ester (II) via the reaction of (I) with dimethyl sulphate in the present of anhydrous of sodium carbonate and dry acetone. Acid hydrazide (III) has been synthesized by adding 80% of hydrazine hydrate to the new ester using ethanol as a solvent. The last step included the preparation of new Schiff-bases (IV-VIII) by the reaction of acid hydrazide with
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