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.
Abstract
The research aims to identify the application of corporate governance requirements according to the international standard (ISO 26000:2010) in the National Insurance Company. Strengths and weaknesses were identified to study the current state of this requirement's application in the company under investigation. The descriptive-analytical approach was utilized through a checklist derived from ISO 26000:2010. Several personal interviews and field visits were conducted to understand the extent of application and documentation based on various statistical methods. The results revealed a level of applic
... Show MoreThe aim of the research is to explain the nature of the relationship between the dimensions of the strategic recovery of the service represented by (compensation, speed of response, apology, initiative (defining the problem) and the strategic goals of the company represented in (profitability, growth, community service, employee satisfaction) in the National Insurance Company, it has been approved The questionnaire as a tool to collect data and information from the sample of (58) who are in (department manager, M. department director, division official, unit official) and the statistical program (spss) was used in calculating (arithmetic mean, standard deviation, coefficient of variation, coefficient of Correlation, t-test, varia
... Show MoreThis study aims to identify the impact of social support on breast cancer patients’ psychological rigidity using a sample in Ramallah and al-Bireh. A descriptive correlative approach was adopted to fulfill the goals of the study and a questionnaire consisted of two criteria: social support and psychological rigidity, which was adopted as a tool for data collection for the study. In order to achieve the goals of the study, the researcher selected a convenient sample that consisted of 123 female breast cancer patients in Ramallah and al-Bireh. This sample represented 50% of the original patient population. The study showed that the average estimated percentage of social support and psychological rigidity for women with breast cancer, in
... Show MoreThis study investigates the impacts of climate change (CC) on the emergence and proliferation of fungal pathogens, with a particular focus on global food security and the potential of medicinal plants and their by-products as sustainable mitigation strategies. Through a systematic literature review of articles published up to 2024, we analyze how CC exacerbates the spread and severity of fungal diseases in crops, leading to significant agricultural losses and threats to food availability. The findings highlight that, alongside conventional approaches such as genetic resistance and precision farming, bioactive compounds derived from medicinal plants and their by-products offer promising, eco-friendly alternatives for the management of fungal
... Show MoreIn addition to their high quantities of active chemicals, medicinal plants are well-known for their pharmacological qualities, which include immunological modulation. T Consequently, this study aimed to examine the effects of Avena sativa and Glycyrrhiza glabra leaf extracts on immunological responses as measured by blood cytokine and liver enzyme levels. The phytochemical analysis of Avena sativa crude leaf extracts revealed the presence of alkaloids,flavonoids, tannins, phenolic compounds, and saponins but the absence of resins and violet oils. On the other hand, violet oils, flavonoids, tannins, saponins, and glycosides were detected in significant concentration in Glycyrrhiza glabra ethanolic extract, although resins and phenolic compou
... Show More
(4)
The widespread use of the Internet of things (IoT) in different aspects of an individual’s life like banking, wireless intelligent devices and smartphones has led to new security and performance challenges under restricted resources. The Elliptic Curve Digital Signature Algorithm (ECDSA) is the most suitable choice for the environments due to the smaller size of the encryption key and changeable security related parameters. However, major performance metrics such as area, power, latency and throughput are still customisable and based on the design requirements of the device.
The present paper puts forward an enhancement for the throughput performance metric by p
... Show More
(3)
(3)
KE Sharquie, AA Noaimi, GA Ibrahim, AS Al-Husseiny, Our Dermatology Online, 2016 - Cited by 3
a laser ablation Q-switched Nd: YAG laser with a wave-length of 355 nm at a variety of laser pulse energies (E) and deposited on porous silicon (PS). Optical emission spectrometer was used to diagnosed medium air to study gold plasma characteristics and prepared Au nanoparticles. The laser pulse energy influence has been studied on the plasma characteristics in air. The data showed the emergence of the ionic (Au II) spectral emission lines in the gold plasma emission spectrum. XRD has been utilized to examine structural characteristics. Moreover, AFM results 37.2 nm as the mean value of the diameter that is coordinated in a shape similar to the rod that appears for Au NPs, in addition to that, TEM has been an indication of the fact that syn
... Show More
(39)
(34)
Background: Hypothyroidism and type 2 diabetes are common conditions worldwide, with hypothyroidism affecting 4–10% of the population and type 2 diabetes affecting approximately 10% of adults globally. The increasing co-occurrence of these two conditions may contribute to increased oxidative stress and associated complications. Low levels of antioxidants are linked to hypothyroidism with type II diabetes because of the growth and production of free radicals, which makes patients more susceptible to oxidative stress. Objective: It was to assess how low antioxidants may exacerbate the pathogenic state of type 2 diabetic hypothyroid patients. Methods: 140 hypothyroid individuals with type 2 diabetes, ages 25 to 60, were assigned to t
... Show More
(4)
(4)