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.
(10)
(2)
The present study was conducted to estimate the incidence, clinical findings, cytological and histopathological characteristics of spontaneously occurring skin neoplasms in dogs. A total of 40 grossly suspected cases of cutaneous and subcutaneous tumors were gathered during the period from July 2016 to August 2018 from male and female dogs in Baghdad city. Dogs with skin neoplasia revealed various clinical signs, and their ages were older than 5 years to 15 years. German shepherd 30% followed by Terrier dogs 25% were more influenced than other breeds. Concerning tumor features, the majority of neoplasms had solitary lesion 70%, regular shapes 65% with black color 55%. The tumors frequently occurred on fore-limbs and abdomen, and 80% of them
... Show More
(5)
(1)
The research aims to identify intelligence spiritual among a sample of students Baghdad University as well as to identify the differences between students in intelligence spiritual according to variable type (male - female), and variable area of study (Science - a human) and variable (First grade - fourth grade), The research sample consisted of (300) students, were applied scale search - a spiritual Intelligence Scale (prepared by the researcher), has resulted in the search results for: -
The students of the University of Baghdad (sample) enjoyed a high level of spiritual intelligence.
- There are no differences between males and females in the spiritual intelligence.
- There
Abstract:
The research aimed to know favoured mass media for children and
modifying their behaviour ,the child became aquires the information from
mass media that he exposure them without any guidance , where upon the
quidance proqrammes becomes real danger whereas qet out their civil
style and converting to deadly poisons,and because of little study for this
supject the two researchers opined to perform astudy to know the favoured
mass media to the children and what are the mass media that modify their
behavior according to ther parent points of view ,after propring the research
measurement and the suilable statical methods it has shown that there are
mass media affect in children behavior ,they are st
This study aims to Statement of the relationship between Total Quality Management philosophy and Organizational performance from the point of view of the internal customer. A comparison has been made between two companies, one of which applies the requirements of TQM well and the other does not apply these requirements as the (General Company for Electrical Industries/ Diyala) and (General Company for Electrical Industries/ Baghdad) to conduct the search, During the questionnaire prepared for this purpose and distributed to a sample of 30 employees in the General Company for Electric Industries/ Diyala and (20) employees of the General Company for Electrical Industries/ Baghdad. Their answers were analyzed using a simple correlation coef
... Show MoreSome types of the fungus Aspergillus were isolated from some hospitals in the city of Baghdad (Imam Ali Hospital and Sadr General Hospital). The samples were taken by Transport media at a rate of three replicates of each place isolated from samples from different places within the hospital (waste, baths, the sick beds, corridors and room floors) for the purpose of isolating and diagnosing the fungus on the Czapeck Dox Agar media. It was noticed that the spread rate of fungus Aspergillus was 70% compared to other species that have emerged during the isolation process of the Sabouraud's Dextrose Agar media. The species A.niger (56.25%) was considered the most common type of fungus visible during the isolation process of the Imam Ali Hospit
... Show MoreStevia rebaudiana Bertoni contains diterpenoid steviol glycosides that have no adverse impact on blood sugar levels despite being 300 times sweeter than sugar. This study aimed to investigate the rate of callus induction from stevia leaves and the content of glycosides when changing the sucrose percentage in the culture medium.. Murashige and Skoog (MS) culture medium supported by 4.0 mg/l naphthalene acetic acid (NAA) and 1.0 mg/l benzyl adenine (BA) was used, and different concentrations of sucrose (2, 3, 4, 5 and 6%) were tested .The extraction of glycosides from leaf and callus tissues was performed by using methanol. Extracted glycosides were analyzed by high-performance liquid chromatography (HPLC). The results showed signific
... Show MoreEM International
The study aims to identify the reality of the use of educational technology in the teaching of mathematics from the point of view of mathematics teachers in Amman, where the researcher used the descriptive survey method. The sample included (67) female teachers working in public schools in Amman. In order to achieve the objectives of the study, the researcher used the questionnaire as a tool to collect data. The study concluded that many educational techniques are not available from the point of view of the study sample. Mathematics teachers also found that there are many obstacles that prevent the use of educational techniques in schools, most notably the lack of courses offered to teachers on the use of teaching techniques in teaching
... Show More