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.
Amino acids have the role in the process of proteins synthesis. They are an essential source of nitrogen atoms that have a role in the pathways of synthetic reaction pathways. The carbon skeletons of the amino acids are the source of energy in addition to their role as precursors in the paths of interactions. The amino acids analysis for the brain of the quail bird in different stage of development (10-16 days of incubation) in addition to the hatching stage (17th day) and the adult. Materials and Methods: Amino Acids Analysis The amino acids were separated from the embryos and adult brains of the quail bird Coturnix coturnix and were diagnosed based on standard amino acids, using high performance liquid chromatographic device (H.P.L.C.). R
... Show MoreExperimental results for the density of states of hydrogenated amorphous silicon due to Jackson et al near the valence and conduction band edges were analyzed using Levenberg-Marquardt nonlinear fitting method. It is found that the density of states of the valence band and the conduction band can be fitted to a simple power law, with a power index 0.60 near the valence band edge, and 0.55 near the conduction band edge. These results indicate a modest but noticeable deviation from the square root law (power index=0.5) which is found in crystalline semiconductors. Analysis of Jackson et al density of states integral J(E) data over about (1.4 eV) of photon energy range, showed a significant fit to a simple power law with a power index of 2.11
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The process of granting loans by banks is the confidence they give to their customers, but this trust should not be a cornerstone in granting loans even if granted these loans on the basis of sound banking should involve risks that may be exposed to the bank because of the failure of the client to meet The bank's financial obligations to the bank due to the unexpected economic conditions affecting the customers, which makes them in a state of faltering, which weaken the ability of banks to provide loans, which are the most important sources of revenue and profits, so the problem of non-performing loans is one of the main problems facing most of the banks Which impede the functioning of its work and the reasons that led to the agg
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Background: Osteoarthritis (OA) currently seems
inevitable and unavoidable for a large swath of the
population .its etiology relates to a strong ,but
complex ,non mendelian genetic basis ,combined
with mechanical and metabolic factors that cause
molecular alterations the end results of which
affect the whole joint .Glucosamine and
chondrotin sulfate alone or in combination may be
of benefit to a subgroup of individuals who have
knee pain due to OA.
Glucosamine has been shown to alter cartiage turn
over in patients with OA undergoing physical
training
Aim of the study: To find the validity of
glucosamine chondrotin sulfate in treatment of
grade 1 and 2 OA.
Methods: the sample of 280 patients
Over the course of six decades, Iraq exposed to many events that have affected the Iraqi people from the social, physical and mental aspects. In this study, two groups of people (2369), from Iraq (G1) and the Michigan, United States (U.S) of America (G2) selected to compare the prevalence rate and effects of trauma factors such as mental illness (anxiety, depression and PTS), somatic diseases (heart disease, hypertension, and diabetes), substances abuse (illicit drugs, alcohol and tobacco), and chemicals pollution), and self-rated health among the two groups. The study results reveals a significant different between the two groups in the all indicators for trauma. The study conclude that Iraqi in U.S. (G2) suffer from factors completely dif
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Introduction: Dental pulp tissue contains many undifferentiated mesenchymal cells, which have the ability to differentiate into different specialized cells. Induced pluripotent stem cells have been developed by various growth factors. The present study was designed to evaluate the effect of application of a combination BMP2/TGF β1 as capped material for traumatic pulp in osteoporotic rat. Materials and Methods: Twelfth female rats (6 normal rat and other 6 osteoporotic rat)., their maxillary anterior teeth subjected to mechanical traumatized pulptomy, the left tooth has speared without treatment, while the right tooth capped with application of 0.5 μl of BMP2 and 0.5μl of TGF β1 .Evaluation of histological changes includes scoring of pu
... Show MoreThe research deals with the problem of visual pollution, as it is one of the most important urban problems that cities suffer from. The concept of visual pollution has recently emerged to describe the deformation and degradation of the urban environment. Visual pollution is defined as any component of the surrounding environment that is inconsistent and not homogeneous with its natural and human components. The volume of visual pollution has doubled due to the non-compliance with the laws, regulations and controls set by the Municipality of Baghdad by the citizens, and to the weak municipal role that the municipality plays in implementing these laws. Therefore, it has become necessary to know the manifestations of visual pollution and th
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The effect of different doses of gamma rays that emitted from 60Co on the development of different stages of lesser grain borer Rhyzopertha dominica (Fab.) was investigated . The results showed that the eggs in both early (1-3 days) and late (7-9) old were very sensitive to gamma rays and its hatch was zero at 10 Gy for both ages in comparison with 83.3% for the control group. Furthermore, the results illustrated that the larval stage in its two old ages were different in the radiosensitivity, the percent of its death were 100% & 96.6% when they exposed to 100 & 120 Gy, respectively. Moreover, the results showed that the sensitivity of pupal stage at age of 1-3 days was increased with increasing the doses of gamma rays, the percent of no
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