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.
Probiotic bacteria using in commercially produced foods are at most members of the genera Lactobacillus and Bifidobacterium. This research was aimed to study the antimicrobial effect of lactobacillus bacteria as probiotic against some pathogenic bacteria, strains of lactobacillus bacteria were isolated from two types of yoghurt in Baquba city, Iraq . Total of 60 yoghurt samples from 2 origins (30 locally and 30 commercial produced yoghurt) were used to isolate probiotic bacteria which were identified phenotypically and biochemically. 30 (100%) of locally produced yoghurt samples include with probiotic bacteria while 12 (40%) commercially types of yoghurt samples were contained with probiotic, the method of modi
... Show More
Aromatic hydrocarbons present in Iraqi national surface water were believed to be raised principally from combustion of various petroleum products, industrial processes and transport output and their precipitation on surface water.
Polycyclic aromatic hydrocarbons (PAHs) were included in the priority pollutant list due to their toxic and carcinogenic nature. The concern about water contamination and the consequent human exposure have encouraged the development of new methods for
PAHs detection and removal.
PAHs, the real contaminants of petroleum matter, were detected in selected sites along Tigris River within Baghdad City in summer and winter time, using Shimadzu high performance liquid chromatography (HPLC) system.
Analysi
From a large number of bacterial samples collected from different hospital in Iraq in central health laboratory ,only ten isolates were identified primary as Vibrio. A number of morphology and biochemical test were carried out to complete this identification that showed all bacterial isolates were related to Vibrio cholerae .In this study all Vibrio isolates were investigated for Bio typing and the result showed that all (10) isolate were related to (Eltor biotypes) .Also, the susceptibility test towards eight antibiotics were carried out .
Results shows that ciprofloxacin , Norfloxacin, Erythromycin, Ampicillin, ceftriaxone and Amikacin were the most effective
... Show More
The origin of occupations and trades producing goods and services necessary for the members of society
When people are designated to perform this task, in which case the state of change is included in the imposition of adequate labor rights, and may change from imposing sufficient to imposing the eye
Does the state have the right to compel "tradesmen and professions" to do their jobs?
Therefore, this research came to address the legitimacy of this from the perspective of the Islamic economy.
Many stone tools were found on a hill south of the Hor Al-Dalmaj which is located in the central part of the alluvial plain of Mesopotamia, between the Tigris and Euphrates Rivers. The types of rocks from which the studied stone tools were made are not found in the alluvial plain, because it consists of friable sand, silt, and clay. All existing sediments were precipitated in riverine environments such as point bar, over bank, and floodplain sediments. The collected stone tools were described with a magnifying glass (10 x) and a polarized microscope after they were thin sectioned. Microscopic analysis showed that these stone tools are made of sedimentary, volcanic igneous and metamorphic rocks, such as: sandstones, limestones, chert, con
... Show More
(1)
This study focuses on diagnosis of Candida species causing Vulvovaginal Candidiasis using phenotype and genotype analyzing methods, and frequencies of candida species also using Vulvovaginal Candidiasis patients. 130 samples (100 from patients and 30 from non infected women) were collected and cultured on biological media. Identifying the yeasts, initially some phenotypic experiments were carried out such as germ tube, from motion of pseudohyphae and clamydospores in CMA+TW80 medium, API20 candida and CHROMagar Candida. Genomic DNA of all species were extracted and analyzed with PCR and subsequent Polymerase Chain Reaction - Restriction Fragments Length Polymorphism (PCR-RFLP) methods. Frequency of C. albicans, C. krusei, C. tropicalis , C.
... Show More
(1)
This article reviews the technical applicability of nanofiltration membrane process for the removal of nickel, lead, and copper ions from industrial wastewater.
Synthetic industrial wastewater samples containing Ni(II), Pb(II), and Cu(II) ions at various concentrations (50, 100, 150 and 200 ppm), under different pressures (1, 2, 3 and 4 bar), temperatures (10, 20, 30 and 40 oC), pH (2, 3, 4, 5 and 5.5), and flow rates (1, 2, 3 and 4 L/hr), were prepared and subjected treated by NF systems in the laboratory. Suitable NF membrane was chosen after testing a number of NF membranes (University of Technology-Baghdad), in terms of production and removal. NF system was capable of removing more than (85%, 78%, and 66% for Ni(II
... 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
... Show More
(1)
Ten isolates of Klebsiella pneumoniae, seven isolates of Pseudomonas aeruginosa and nine isolates of Staphylococcus aureus, were obtained from 100 urine samples collected from Baghdad hospitals. All isolates were identified biochemically and confirmed by using VITEK 2 and were then tested for their susceptibility towards 6 antibiotics and for phenolic extracts of Thymus vulgaris and Cinnamomum cassia. All bacteria were greatly affected by T. vulgaris, especially K. pneumoniae. Viable count was performed, it was noted that the number of bacterial cells reduced from 1×108 CFU to 1.2× 103, 2×105 and 1.8×106CFU of K. pneumoniae, P. aeruginosa and S. aureus respectively. While C. cassiahad a slight effect on them. K. pneumoniae isola
... Show More
The aim of this paper was to investigate the removal efficiencies of Zn+2 ions from wastewater by adsorption (using tobacco leaves) and forward osmosis (using cellulose triacetate (CTA) membrane). Various experimental parameters were investigated in adsorption experiment such as: effect of pH (3 - 7), contact time (0 - 220) min, solute concentration (10 - 100) mg/l, and adsorbent dose (0.2 - 5)g. Whereas for forward osmosis the operating parameters studied were: draw solution concentration (10 - 150) g/l, pH of feed solution (4 - 7), feed solution concentration (10 - 100) mg/l. The result showed that the removal efficiency by using adsorption was 70% and the removal efficiency by using forward osmosis was 96.2 %.
... Show More