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.
First: the definition searchEnsure the definition Introduction of research and its importance, as it was addressed to the importance of the importance of training in the development of people and institutions and the importance of diversifying the training vocabulary and a lot of relevant courses for its role to develop work and get away from the routine work, and was put up for the importance of research to identify the problems and obstacles facing the process of providing training as the best in order to access to the ambition sought by workers on training and even the trainees while defined research problem that was observed weak performance and cost-effective training through the study of some of the questionnaires distributed
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The study included the collection of samples of raw cow milk to isolate Leuconostoc bacteria, samples were sub cultured on De-Man Rogosa Sharpe-Vancomycin medium, the pure colonies were selected and subjected to the cultural and microscopically tests, according to that 25 cocci bacterial isolates were obtained, then isolates were subjected to biochemical tests. Result of tests showed that 12 isolates belong to the genus Leuconostoc out of 25 cocci bacterial isolates, Vitek2 system was used as a supplementary step. Results of final identification showed that 3 sub species were obtained included Leuconostoc mesenteroides ssp. cremoris 9 out of 12 isolates, while it was 2 isolates of Leuconostoc mesenteroides ssp. mesenteroides and one isol
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Time is very important in educational institutions. It is also one of our contemporary problem ‚as time is a clear – cut and limited factor‚ it demands that administrators should monitor it by administering and monitoring the principles of time.
Hence‚ the researcher attempts to identify the skills of administrating time and the reasons that cause the waste of time of the Heads of Departments at university of Baghdad.
Significance of the research:
Time is very important to all educational administrators and one of them is the institutions of Higher education. One of the
... Show MoreThis study aims to investigate the academic leaders’ perceptions towards the degree of availability of the dimensions of organizational immunity systems at the University of Tabuk, as well as to reveal the statistically significant differences between the average responses of the study sample members about the degree of availability of these dimensions at their university due to the variables of (gender, leadership position, and college specialization). To achieve the objectives of the study, a descriptive survey method was used. The study population consisted of (200) male and female leaders who were academic leaders at the University of Tabuk. A questionnaire was used as a tool for collecting data, which its validity and reliability
... Show MoreThe goal of current research is to identify the difficulties in the application of modern physics in the middle schools of the province of Baghdad schools from the perspective of teachers of physics trends, sample search of (127) teachers, Karkh Third Directorate, and use Researcher questionnaire data collection tool after applying it to teachers who have experience (5) years and more after confirmation of the validity and reliability of the scale (the tool) researcher has used the averages for the purpose of interpreting the results. the results showed that the difficulties have been in the order following: (difficulties related educational environment of modern trends of teaching, curriculum-related dif
... Show MoreThis work deals with thermal cracking of heavy vacuum gas oil which produced from the top of vacuum distillation unit at Al- DURA refinery, by continuous process. An experimental laboratory plant scale was constructed in laboratories of chemical engineering department, Al-Nahrain University and Baghdad University. The thermal cracking process was carried out at temperature ranges between 460-560oC and atmospheric pressure with liquid hourly space velocity (LHSV) equal to 15hr-1.The liquid product from thermal cracking unit was distilled by atmospheric distillation device according to ASTM D-86 in order to achieve two fractions, below 220oC as a gasoline fraction and above 220oC as light cycle o
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