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 study was done to find a cheap, available and ecofriendly materials that can remove eosin y dye from aqueous solutions by adsorption in this study, two adsorbent materials were used, the shells of fresh water clam (Cabicula fluminea) and walnut shells. To make a comparison between the two adsorbents, five experiments were conducted. First, the effects of the contact time, here the nut shell removed the dye quickly, while the C. flumina need more contact time to remove the dye. Second, the effects of adsorbent weight were examined. The nut shell was very promising and for all used adsorbent weight, the R% ranged from 94.87 to 99.29. However C. fluminea was less effective in removing the dye with R% ranged from 47.59 to 55.39. The thi
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Eimeriosis is a major problem affecting ruminants worldwide. The disease is primarily caused by Eimeria species, which are specialized for each host and grow in the small and large intestine of animals. The losses due to subclinical infections (especially weight loss) and clinical disease (diarrhea) make the species of this genus a very significant economic concern. Therefore, this study was conducted in some areas of Wasit Province. A total of 180 fecal samples from goats, of both sexes and covering different age groups and months, were collected. All fecal samples were examined microscopically, and 75 positive fecal samples were taken for molecular examination and further analyzed using conventional PCR, sequencing and phylogeneti
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In this work, the detection of zinc (Zn) ions that cause water pollution is studied using the CSNPs- Linker-alkaloids compound that was prepared by linking extracted alkaloids from Iraqi Catharanthus roseus plant with Chitosan nanoparticles (CSNPs) using maleic anhydride. This compound is characterized by an X-ray diffractometer (XRD) which shows that it has an orthorhombic structure with crystallite size in the nano dimension. Zeta Potential results show that the CSNPs-Linker-alkaloids carried a positive charge of 54.4 mV, which means it possesses high stability. The Fourier transform infrared spectroscopy (FTIR) shows a new distinct band at 1708.93 cm-1 due to C=O esterification. Scanning electron microscope (SEM) image
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For this research, the utilisation of electrocoagulation (EC) toremove theciprofloxacin (CIP) and levofloxacin (LVX) from aqueous solutions was examined. The effective removal efficiencies are 93.47% for CIP and 88.00% for LVX, under optimum conditions. The adsorption isotherm models with suitable mechanisms were applied to determine the elimination of CIP and LVX utilizingtheEC method. Thefindingsshowed the adsorption of CIP and LVX on iron hydroxide flocs followed the Sips isotherm, with correlation coefficient values (R2) of 0.939 and 0.937. Threekinetic models were reviewed to determine the accurate CIP and LVX elimination methods using the EC method. The results showed that itfittedfor the second-order model, which indicated that the c
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The aim of this work is to detect the best operating conditions that effect on the removal of Cu2+, Zn2+, and Ni2+ ions from aqueous solution using date pits in the batch adsorption experiments. The results have shown that the Al-zahdi Iraqi date pits demonstrated more efficient at certain values of operating conditions of adsorbent doses of 0.12 g/ml of aqueous solution, adsorption time 72 h, pH solution 5.5 ±0.2, shaking speed 300 rpm, and smallest adsorbent particle size needed for removal of metals. At the same time the particle size of date pits has a little effect on the adsorption at low initial concentration of heavy metals. The adsorption of metals increases with increas
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The shape dimensions and characteristics of pollen grains and seeds have importance in distinguish among species. Therefore, the present study included morphological characteristics of pollen grains and seeds for eight species belonging to eight genera of the family Brassicaceae and these species are: Alliaria petiolata (M.Bieb) Cavara et Grand, Aubrieta parviflora Boiss, Cardamine hirsuta L., Crambe orientalis L., Eromobium aegyptiacum (Spreng.) Schweinf.et Asch.ex Boiss., Parlatoria cakiloidea Boiss., Sterigmostemum sulphureum (Banksetsol.) Bornm. Neotorularia torulosa (Desf.) Hedge & J. Leonard. The pollen grains were studied in morphological and full measurements were taken, the study showed that the majority of the pollen grai
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