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 present study provides the first record species of the genus Lithobius Leach, 1814, L. ferganensis (Trotzina, 1894) which was collected from the middle of Iraq. A detailed explanation of the morphology and the diagnostic characters of specimens of both sexes is provided.
The kinetic of atropine pertraction from seeds of Datura Metel Linn plant was studied. Diisopropyl ether, n-hexane and n-heptane were used as membranes for atropine recovery. The effect of speed of agitation and time in the range of 200-300 rpm and 0-3.5h, respectively were studied using the proposed membranes. The pertraction experiments were carried outs in a batch laboratory unit. The liquid-liquid pertraction was found to be very suitable for atropine recovery from its liquid extracts of Datura Metel seeds. A high purity (94-96%) can be obtained in the receiver phase. The pertraction process was found to be very selective for atropine recovery with diisopropyl ether membrane. As the speed of agitation increases the efficiency of pertrac
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Education specialists have differed about determining the best ways to detect the
talented. Since the appearance of the mental and psychological measurement movement, some
scholars adopted intelligence ratios as a criterion to identify the talented and others went to
rely on the degree of academic achievement. Each of these two methods has its own flaws and
mistakes and a large number of talented children were victims of these two methods.
Therefore the need to use other scales for the purpose of detection of talented children
appeared because they provide valuable information which may not be obtained easily
through objective tests and these scales are derived from consecutive studies of gifted andtalented children
In this study, synthesised new ligand: potassium 2,2'-(quinoxaline-2,3- diyl)bis(1-phenylhydrazinecarbodithioate) (L). The ligand synthesised by reacting N1,N2-dip-tolyloxalamide as the starting material with CS2 and KOH to add the CS2 group and then with phenylendiammine to achieve (L). The ligand used in the synthesis of complexes with (CoII, NiII and CdII). The new ligand and its complexes characterised by FT-IR, UV-Vis, 1H, 13C-NMR, Mass spectroscopy, and elemental analysis, in addition to the above techniques were using magnetic moment, atomic absorption, chloride content, and melting point to describe the metal complexes.
This work investigates the effect of the gas nitriding process on the surface layer microstructure and mechanical properties for steel 37, tool steel X155CrVMo12-1 and stainless steel 316L. Nitriding was conducted at a temperature of 550 °C for 2 hours during the first stage and at 750 °C for 4 hours during the second stage. SEM and X-ray diffraction tests were performed to evaluate the microstructural features and the major phases formed after surface treatment. SEM and X-ray diffraction tests were performed to assess the microstructural features and the primary phases formed after surface treatment. The new secondary precipitates were identified as γ′-Fe4N, ε (Fe2–3N), and α-Fe, exhibiting an uneven chain-like pattern wit
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The lesser grain boner Rhizopertha dominica is an important insect on grain in Iraq . in this study phenol and coumarin were isolated from Albizzia lebbek . the addition of there compounds to artificial diet at concentration of 3.4,5% couse high effects on the development of the studed insect larva.the larva were unable to complete its development to adult stage. More over, using 5% coumarin larva were died.Seed germination was not affected when seed were sprayed with concentration of 3,4,5% of coumarin and phenol.
The present study introduces description of a new species of genus Arboridia Zakhvaticin 1946, based on a large collection of Cicadellids. External morphological characters particularly male genitalia were discussed and illustrated. The genus Arboridia Zalchvatkiia (Typhlocybinae: Erythroneurini) contains small slender, fragil and attractively coloured and patterned leafhoppers. It was erected by Zakhvatkin in 1946 (Zalchvatkin, 1946). The overall length of adults ranges from 2.5 to 3.4 mm. Members of this genus can be recognized by inner apical cell of forewing which is long with oblique base; Cu confluent with this base at a point near the middle of the length of inner apical cell; two prominent circular deep brown spots on vertex (Zal
... Show MoreThe purified frog skin peptides were tested on leukemic patients lymphocytes, which revealed effects of cytotoxicity. Four frogs (Rana ridibunda) were stimulated by single intra-peritoneal injection of norepinephrine-HCl . Five different peptides;1(18) A, 2(19) L, 3(20) I,4(21) E and 5(22) Y were isolated and quantified. The peptide 3(20)I had 5.87% of hemolysis, while healthy human lymphocytes cytotoxic activity was for 2(19)L with inhibition( -10.4%).All peptides were subjected to polyacrylamide gel electrophoresis. The results revealed peptides 1(18)A, 2(19)L, 3(20)I which appeared as low as 10 KDa marker. Theoretically, the whole polypeptide had a molecular weight 7488.61 Dalt
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The present study was conducted to estimate the incidence, clinical findings, cytological and histopathological characteristics of spontaneously occurring skin neoplasms in dogs. A total of 40 grossly suspected cases of cutaneous and subcutaneous tumors were gathered during the period from July 2016 to August 2018 from male and female dogs in Baghdad city. Dogs with skin neoplasia revealed various clinical signs, and their ages were older than 5 years to 15 years. German shepherd 30% followed by Terrier dogs 25% were more influenced than other breeds. Concerning tumor features, the majority of neoplasms had solitary lesion 70%, regular shapes 65% with black color 55%. The tumors frequently occurred on fore-limbs and abdomen, and 80% of them
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