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.
Shear wave velocity is an important feature in the seismic exploration that could be utilized in reservoir development strategy and characterization. Its vital applications in petrophysics, seismic, and geomechanics to predict rock elastic and inelastic properties are essential elements of good stability and fracturing orientation, identification of matrix mineral and gas-bearing formations. However, the shear wave velocity that is usually obtained from core analysis which is an expensive and time-consuming process and dipole sonic imager tool is not commonly available in all wells. In this study, a statistical method is presented to predict shear wave velocity from wireline log data. The model concentrated to predict shear wave velocity fr
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Abstract: Coronavirus disease 2019 (COVID-19) is an infectious disease with severe acute respiratory syndrome and first recognized in Wuhan, China, and it has since spread to the world, resulting in the coronavirus pandemic to 2020. The present study aimed to evaluate Molecular study of some types of vaginal fungi isolated from recovered women from Covid-19 in Baghdad governorate. The study was conducted on 213 samples collected between December 2021 and March 2022, where the number of positive samples reached 188 with percentage 88.26%, while the number of negative samples reached 25 with percentage 11.73% by taking vaginal swabs from various female patients in Al- Kadhimiya Teaching Hospital. Three of Candida spp. were isolated: Candida a
... Show MoreP. aeruginosa is one of the complex targets for antimicrobial chemotherapy. Also, it is intrinsically resistant to several antibiotics. It produces β-lactamases enzymes that are responsible for the widespread β-lactam antimicrobial resistance. There are three major groups of β-lactamase enzymes, MBLs and ESBLs forming Pseudomonas is a major issue for the treatment of burns victims. Methods: A total of 28 clinical isolates related to P. aeruginosa have been obtained from the burns specimens from patients attending to AL-Imam hospital/Baghdad-Iraq, through the period from October 2015 to March 2016. Also, all isolates have been recognized as P. aeruginosa via utilizing bacteriological assay and confirmed by Vitek 2. In addition, the suscep
... Show MoreA rapid and sensitive method for analysis of amino acid hydrolysates of nigella sativa L seed has been developed using O-phthaldialehyde(OPA ) as a pre-column derivatizing agent. OPA reagents in the presence of mercaptoethanol react rapidly with primary amino acids ( less than 60 sec.) to form isindole derivatives which easily separated with good selectivity on ODS column. Resolution of amino acid derivatives is carried out with a methanol gradient in 0.01 maqueous sodium acetate. pH 7.1 . The quantitation of amino acid derivatives is reproducible within an average relative deviation of + 1.4% the linearity for most amino acids were more than 0.9993 with detection limit of 0.2 ppm. 15 amino acid were detected in the analysis of
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Biological activity substances was investigated in watery extract of lentil which found to contain phenols, tannin, saponins and resins while, flavons, terpens and steroids were not exist in the extract details explained that 5%, 10% of lentil extract largly inhibited the growth of Psedumonas aeruginosa then Escherichia coli and Bacillus subtilis. The growth of both Staphylococcus aureus and Salmonella typhimurium were slightly affected by all extract concentration. Extracellular protease were screened in all bacterial species under study. Complete inhibition was achieved for extracellular protease while different percentage of protease inhibition were seen for intracellular proteases.
ABSTRACT
Two compounds were isolated from the fruit part of Rhus coriaria that grow wildly or cultivated in the north of Iraq. The compounds were separated by preparative high-Performance Liquid Chromatography and their structures were established based on detailed spectroscopic techniques like FTIR and LC-MS/MS.
Keywords: Rhus coriaria, Preparative HPLC, LC-MSMS, FTIR
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This study aimed at isolating uropathogenic Escherichia coli from urinary tract infections (UTIs) of human and cattle to examine the molecular diversity and phylogenetic relationship of the isolates. A total of 100 urine samples were collected from UTIs of human and cattle. The isolates identification was done using routine diagnostic methods and confirmed by Vitek2. Antimicrobial susceptibility was tested against 10 antimicrobials. Random amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR) was applied to identify the genetic diversity among E. coli isolates from human and animal origin by using five different octamer primers. The gelJ software for the phylogenetic analysis created Dendrograms. Out of 50 human urine samples, E.
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The foreguts of a total of 515 fish of Chondrostoma regium (Heckel, 1843) (locally: Bala’aot Malloky) were studied. These fish were collected from Tigris River at Salah Al-Deen Province (between Al-Hagag & Yathrib) for 20 months between March and October of the next year. Detritus, plant in origin materials (19.6%, 23.0% & 24.9%); green and blue green algae, mostly Cladophora, Cosmarium and Merismpedia sp. (17.1%, 12.9% & 12.2%) and diatoms, mostly Diatoma, Chanathes, Amphora and Cyulbella sp. (16.9%, 8.8% & 8.2%) were the main food categories taken by these fishes according to occurrence (O%), volumetric methods (V%) and ranking index (R%). Debris (not part of the diet) took 45.3% of the studied fish foreguts by volume. Detritus was also
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In this paper, the species of the genus of Chlaenius Bonelli, 1810 (Coleoptera, Carabidae) were reviewed, and it was revealed that there are 21 confirmed species in Iraq; among them, the species of Chlaenius hamifer Chaudoir, 1856 was recorded for the first time in Iraq.
Diagnostic characters, a redescription of some of the morphological features, photographs and illustrations are provided for the new record species in this investigation.