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.
(1)
(1)
This work was carried out to investigate the shrinkage properties of insulator ceramic body, and the effect of changing the ratio of concentration of electrolyte solution on these properties. The electrical insulator body was prepared by using Iraqi local materials. These are :- kaolin Duakhla, Arudhuma Sand glass, and potash feldspar with weight percentage (45%, 25%,and 30%) respectively. The processes of milling, classification, mixing, forming and firing, through the process of mixing electrolyte solution and mineralizer were added at different concentrations for electrolyte solution, (1%, 0.7%, 0.5%, 0.2%, and 0.1%). The electrolyte solution was prepared by mixing sodium carbonate and sodium silicate by (2:1) ratio
... Show MoreImplementing smart community engagement should consider careful planning and collaboration with numerous stakeholders, including the community. The technology and program must be designed to frame its purpose and should link back to specific goals of implementing smart community engagement. Digital services do not guarantee a smart engagement between the community and the local government. This is the case for the Kubang Pasu local government where several online services have been provided in their attempt to implement the smart community concept. However, understanding on the preferences of features and requirements of existing web-based systems and the impact of these systems is lacking. Therefore, a perception study needs to be condu
... Show More
(4)
(3)
Abstract :
This research aims to examine the correlation and the impact of Cultural Intelligence with their dimensions (Strategy, Knowledge, Motivation, Behavior) on Drawing Local politics and their dimensions (Empowerment, Flexibility, Organizational Justice, Local Funding) In Dhi Qar Provincial Council To determine the extent of the presence of significant statistical differences between research variables Due to the recent experiment which requires clarification of the role of the pivotal and important carried out by the provincial council in the exercise of his work in light of the diversity of cultures and the peculiarities of the local community, which may impede the provision of equal services to all those parties
... Show More
Abortion is a condition that occurs due to one of the pathological injuries, often one of the members of the TORCH is the real cause. The current study aimed to investigate the impact of toxoplasmosis, HSV-2 infections with abortion, and also, the identification of immunogenetics marker (interleukin-10) that may be associated with abortion. Anti-Toxoplasma IgG, IgM, Herpes simplex virus-2 IgM, human soluble leukocyte antigen class I–G and interleukin10 were estimated by ELISA technique, while the expression of IL-10 gene was investigated by using the real-time PCR. The results showed that among aborted women the rate of anti-Toxoplasma and HSV-2 IgM antibodies occurred within the age groups (21–30) years and (31–40) years 32(100.0%) a
... Show More
(21)
In the drilling and production operations, the effectiveness of cementing jobs is crucial for efficient progress. The compressive strength of oil well cement is a key characteristic that reflects its ability to withstand forceful conditions over time. This study evaluates and improves the compressive strength and thickening time of Iraqi oil well cement class G from Babylon cement factory using two types of additives (Nano Alumina and Synthetic Fiber) to comply with the American Petroleum Institute (API) specifications. The additives were used in different proportions, and a set of samples was prepared under different conditions. Compressive strength and thickening time measurements were taken under different conditions. The amoun
... Show More
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
... Show More
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
... Show More
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 More