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.
Background: The gene responsible for encoding the protein of cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) has been found to be associated with rheumatoid arthritis (RA) in different ethnic populations. But the association of +49A/G CTLA-4 polymorphism with susceptibility of RA among Iraqi Arab populations has not yet been determined. Methods: One hundred and seventy-eight patients were examined, 67 of them were males (mean age 54.71 ± 10.4 years), while 167 were examined for the control group, of whom 64 were males and the rest were females. CTLA-4 DNA genotyping was carried on to determine the +49 A/G (rs231775) polymorphism using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Enzyme-linked immuno
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A method was developed that offers a rapid, simple and accurate technique for the determination of chlorophenols at trace levels in aqueous samples with very limited volumes of organic solvents. These compounds were acetylated, then preliminarily extracted with n-hexane. The enriched chlorophenols were directly analyzed using gas chromatography with an electron-capture detector. The detection limits were in the range of 0.001–0.005 mg/L, except for 2-chlorophenol, which was always above 0.013 mg/L. Relative standard deviation for the spiked water samples ranged from 2.2 to 6.1%, while relative recoveries were in the range of 67.1 to 101.3%.
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In this paper we prove the boundedness of the solutions and their derivatives of the second order ordinary differential equation x ?+f(x) x ?+g(x)=u(t), under certain conditions on f,g and u. Our results are generalization of those given in [1].
This study was conducted to evaluate the efficacy of different techniques for extraction and purification of Tomato yellow leaf curl virus (TYLCV). An isolate of the virus free of possible contamination with other viruses infecting the same host and transmitted by the same vector Bemisia tabaci Genn. was obtained. This was realized by indicator plants and incubation period in the vector. Results obtained revealed that the virus infect Nicotiana glutinosa without visible symptoms, while Nicotiana tabaccum var. White Burley was not susceptible to the virus. The incubation period of the virus in the vector was found to be 21 hrs. These results indicate that the virus is TYLCV. Results showed that Butanol was more effective in clarification the
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Leucine aminopepotidase (LAP)[EC:3.4.11.1] activity has been assayed in (50) serum samples of patients with diabeties naphrophathy D.N (non-insulin dependent diabetic (NIDD) , and (50)serum sample of healthy individuals without any clinically detectable diseases have been as control group. The aim of this study is to measure leucine aminopeptidase activity and partially purifying the enzyme from sera of patients with diabetes nephropathy The results of this study revealed that Leucine aminopeptidase (LAP) activity of nephropathy patient’s serum shows a high signifiacant increase (p < 0.001) compared to that of the healthy subjects.LAP was purified from the serum of patients with diabetes nephropathy by dialysis and gel filtration (Se
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This study aimed to determine the optimal conditions for extracting basil seed gum in addition to determine the chemical components of basil seeds. Additionally, the study aimed to investigate the effect of the mixing ratio of gum to ethanol when deposited on the basis of the gum yield which was1:1, 1:2, 1:3 (v/v) respectively. The best mixing ratio was one size of gum to two sizes of ethanol, which recorded the highest yield. Based on the earlier, the optimal conditions for extracting basil seed gum in different levels which included pH, temperature, mixing ratio seeds: water and the soaking duration were studied. The optimal conditions were: pH 8, temperature of 60°C, mixing ratio seeds: water 1:65 (w/v) and soaking duration of 30 min
... Show MoreThe investment budget represents a stage of the investment decision in service units, and the preparation and implementation needs to be a complement of the same planning part, because the planning does not end with the development of the plan, but includes a follow-up implementation, so it has to be effective and efficient oversight of the estimates and procedures for disbursement of funds approved for investment projects, The problem with research in that local governments suffer from the presence of Allkaat and problems facing the implementation of the investment budget projects due to the adoption budget items which can not be measured the efficiency of the performance of these units of government by, and shortcomings in the control
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Sera samples were collected from 60 children aged 4-60 months, all were clinically and serologically proven cases of visceral leishmaniasis, as well as from 10 healthy children, all were seronegative with no history of parasitic infection who serve as a control during the study. Serum total protein and albumin were measured and compared between the control and visceral leishmaniasis patients. Serum protein profiles have been investigated using the conventional sodium dodecyl sulphate – polyacrylamide gel electrophoresis (SDS-PAGE). Serum of control group showed the specific protein pattern with five protein bands, while serum protein profile in visceral leishmaniasis pat
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Molasse medium containing different concentrations of (NH4)2 SO4, (NH4)3 PO4, urea, KCI, and P2O5 were compared with the medium used for commercial production of C. utilis in a factory south of Iraq. An efficient medium, which produced 19. 16% dry wt. and 5. 78% protein, was developed. The effect of adding various concentrations of micronutrients (FeSO4, 7T20, MnSO4. 7H20, ZnSO4. 7E20) was also studied. Results showed that FeSo4. 7H20 caused a noticeable increase in both dry wt. and protein content of the yeast.