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.
In this research study theory to find the stress and emotion gases in the glass as a result of exposure to pulses of the laser beam has been the study using vehicles three major on-system axes cylindrical (r, 0, z), where I took three models of glass silica glass soda glass fused and shedtwo types of lasers where the study showed that the thermal stresses and emotions ...
Abstract
The current research aims to clarify the role of local policies on the performance of the province of Baghdad, after studies proved practical experience what those policies of the major role and effect on the lives of citizens, as well as alleviate the burden on central government, which make a lot of states give local governments broad powers and her specialty funds for the exercise of its vital role and actor in various joints of local development, research has indentified a problem in a number of questions such as: do you have the policy of the provincial council of local qualified and able to influence the performance of the province? What are the main forces of powerful and implementation of policies at all
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This paper study the ability of Enterobacter cloacae for degrading crude oil in contaminated water. Six isolates of E. cloacae were isolated from hydrocarbon contaminated soil and water of different sites. The isolate E. cloacae E1 showed the highest emulsification index (E24%) reached 62% thus it was chosen for further study. Biosurfactant produced by E. cloacae E1 reduced the surface tension of the medium from 64 to 36 mN/m. pH range 6.5 – 7 and temperature range 30˚C - 35˚C were the optimal conditions for maximum degradation. After 30 days of incubation, E. cloacae E1 degraded 70.00 ± 0.40% of the crude oil. GC-MS analysis revealed that E. cloacae E1 was able to degrade aromatic compounds. This study proved that E. cloacae E1 con
... Show MoreDermatophytes are species with slight genetic variation, and are yet several uncertainties about the differences among species. This study aims to isolate and diagnose the Trichophyton interdigitale by molecular technique and to reveal the phylogenetic distance and similarity of the Iraqi isolates to other isolates from the globe, in addition, to submit the obtained sequences to the NCBI database. This study included 86 with multiple lesions on different parts of the body. The results showed different variations within the ITS gene between the isolates. It was concluded that Trichophyton interdigitale in Iraqi isolates had two types of substitution variations (Transition and Transversion) different than global isolates. Moreover, it
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Traffic loading and environmental factors are among the most serious variables that cause the spoilage of flexible pavements and lead to a decrease in their design life. The objective of this study is to investigate the influence of axle load raise and the change in resilient modulus on the flexible pavement design life. Locally, Highway geometric design code for Iraqi building code has assign certain admissible maximum load limits per every axle truck type that should not be overrun. In this paper nine different axle truck loads (8, 9, 10, 11, 12, 13, 14, 15, and 16) tons, single axle with dual tire and, and two different resilient moduli of asphalt pavement were chosen. The evaluation was carried out assuming high temperature to represent
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According to the prevalence of multidrug resistance bacteria, especially Pseudomonas aeruginosa, in which the essential mechanism of drug resistance is the ability to possess an efflux pump by which extrusion of antimicrobial agents usually occurs, this study aims to detect the presence of mexB multidrug efflux gene in some local isolates of this bacteria that show resistance towards three antibiotics, out of five. Sensitivity test to antibiotics was performed on all isolates by using meropenem (10µg/disc), imipenem (10µg/disc), amikacin (30 μg/disc), ciprofloxacin (5µg/disc) and ceftazidime (30 µg/disc). Conventional PCR results showed the presence of mexB gene (244bp) in four isolates out of t
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According to the prevalence of multidrug resistance bacteria, especially Pseudomonas aeruginosa, in which the essential mechanism of drug resistance is the ability to possess an efflux pump by which extrusion of antimicrobial agents usually occurs, this study aims to detect the presence of mexB multidrug efflux gene in some local isolates of this bacteria that show resistance towards three antibiotics, out of five. Sensitivity test to antibiotics was performed on all isolates by using meropenem (10μg/disc), imipenem (10μg/disc), amikacin (30 μg/disc), ciprofloxacin (5μg/disc) and ceftazidime (30 μg/disc). Conventional PCR results showed the presence of mexB gene (244bp) in four isolates out of ten (40%). In addition,25, 50μg/ml of cur
... Show MoreScience occupies great importance in Islamic thought. Science and learning are considered an essential part of Islamic teachings, and this importance appears in several aspects,Among them is thatScience as a means of understanding religion :Science is a means of understanding the teachings of the Islamic religion. Islam encourages thinking and rational research to understand the Holy Quran and the Sunnah of the Prophet, enabling Muslims to direct their lives and actions in accordance with the directives of their religion,And also to encourageResearch :Islam encourages scientific research and the use of reason in understanding the nature of the universe and God’s signs in it. Muslims are encouraged to study the natural and social s
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This studies p- CuO / n - Si hete-rojunction was deposited by high vacuum thermal evaporation of Copper subjected to thermal oxidation at 300 oC on silicon. Surface morphology properties of The optical properties concerning the transmission spectra were studies for prepared thin films. this structure have been studied. XRD anaylsis discover that the peak at (𝟏𝟏𝟏-) and (111) plane are take over for the crystal quality of the CuO films. The band gap of CuO films is found to be 1.54 eV. The average grain size of is measured from AFM analysis is around 14.70 nm. The responsivity photodetector after deposited CuO appear increasing in response