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.
(31)
(20)
Silver nanoparticles synthesized by different species
Abstract Ternary Silver Indium selenide Sulfur AgInSe1.8S0.2 in pure form and with a 0.2 ratio of Sulfur were fabricated via thermal evaporation under vacuum 3*10-6 torr on glasses substrates with a thickness of (550) nm. These films were investigated to understand their structural, optical, and Hall Characteristics. X-ray diffraction analysis was employed to examine the impact of varying Sulfur ratios on the structural properties. The results revealed that the AgInSe1.8S0.2 thin films in their pure form and with a 0.2 Sulfur ratio, both at room temperature and after annealing at 500 K, exhibited a polycrystalline nature with a tetragonal structure and a predominant orientation along the (112) plane, indicating an enhanced de
... Show More
The Necessity of College of Education for Woman from the Point of view of Students
This study aimed to conduct a comparative analysis of seven calliphorid species of third- instar larvae that were collected from central Iraq. The purpose was to morphologically characterize these species and develop an identification key to differentiate them from closely related species. Scanning Electron Microscope (SEM) graphical images and high-resolution traditional optical microscopes were used to analyze the morphological characteristics of the species Calliphora vomitoria (Linnaeus, 1758), C. vicina Robineau-Desvoidy, 1830, Chrysomya albiceps (Widemann, 1819), Ch. rufifacies (Macquart, 1844), Ch. megacephala (Fabricius, 1794), Lucilia cuprina (Wiedemann, 1830), and L. sericata (Meigen, 1826). An identification key was const
... Show More
(1)
(1)
ABSTRACT In dam construction stages when an earth embankment has retained a reservoir with constant water surface elevation for a long time, seepage conditions within the embankment will be reach a steady state. If it is necessary to drain the reservoir quickly, the pore-water pressures in the embankment may remain relatively high while the stabling effect of the reservoir's weight along the upstream (U/S) side for the embankment has removed. This process is referring to as "Rapid Drawdown" and may be cause instability in the upstream (U/S) face of the embankment. Kongele dam is one of the proposed earth dams to be implement within the current plan in Iraq. The authors study pore water pressure and the effect of rapid drawdown for the dam d
... Show MoreThree isolated bacteria were examined to remove heavy metals from the industrial wastewater of the Diala State Company of Electrical Industries, Diyala-Iraq. The isolated bacteria were identified as Pseudomonas aeruginosa, Escherichia coli and Sulfate Reducing Bacteria (SRB). The three isolates were used as an adsorption factor for different concentrations of Lead and Copper (100, 150, and 200 ppm.), in order to examine the adsorption efficiency of these isolates. In addition, the effect of three factors on heavy metals adsorption were examined; temperature (25, 30, and 37 ?C), pH (3 and 4.5) and contact time (2 and 24 hrs). The results showed that the highest level of lead adsorption was obtained at 37 ?C by E. coli, P, aerugenosa and
... Show More
(5)
The fauna of bees (Hymenoptera, Apoidea) from different regions of Iraq is surveyed in this study; there were 16 species, 13 genera that belong to four families which are collected in this investigation.
Also, all the species that are recorded for Iraq in previous investigations are revised; totally there are 110 species, 32 genera belonging to five families: Apidae, Andernidae, Colletidae, Halictidae and Megachilidae were listed.
(15)
(8)
This work includes a detailed description of the Leucostoma nigricorpuris sp. nov. from
Iraq. Locality, host plants and data of collection were given.