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.
This study was conducted to provide a detailed description of the osteology features of Alburnus amirkabiri from the Qareh Chai river, markazi province, Iran. For this purpose, eight specimens of A. amirkabiri were collected from the Qareh Chai River by electrofishing and fixed in 4% buffered formalin after anesthesia. The specimens were cleared and stained for osteological examination and its detailed osteological characterizations and differences with available osteological data of other members of the genus Alburnus were provided.
Influential, organized groups with natural antimicrobial and anti-biofilm broad-spectrum power exist within the food chain, like a hidden dormant mimic hygienic bio life nanobodies that can terminate multiple opportunistic disease entities owing multi-stress resistant forbidden recalcitrant power, such as Candida albicans. These wonderful dynamic forces created by ALLAH Almighty are the Mycophages or fungi-eating state of fungi foodborne phages, and this project was redirected to be a dare to leap from us towards the future. Multi-stress resistant C. albicans that are resistant to different antifungal agents with their genetic tolerance plasticity to thermal pasteurization decontamination module as well as to ultraviolet irradiation
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Background: The miswak is a teeth cleaning twig made from a twig of the Salvadora persica tree (known as arak in Arabic).A traditional alternative to the modern toothbrush, it has a long, well-documented history and is reputed for its medicinal benefits. It also features prominently in Islamic hygienical jurisprudence. Materials and methods: Twenty maxillary first premolars were treated with the selected solutions which included siwak water extract (5%, 10%) and sodium fluoride 0.05% for 2minutes once daily for 20 days interval, deionized water was used as control negative. Then the concentration of the dissolved calcium ion in the etching solution of 2N HCL was measured. Results: The least amount of the dissolved calcium ion was registere
... Show MoreCyprinidae species are the most abundant and widely distributed fish species in the inland waters of Iraq. Cyprinids are complex species, and it is difficult to identify them on the basis of morphology. Thus, the morphological characteristics must be achieved and confirmed by molecular analysis. Twenty specimens of Cyprinion kais Heckel, 1843 (Piscies, Cypriniformes, Cyprinidae) were collected from two localities at Tigris River in the middle of Iraq: five specimens from Al-Tharthar Lake, Saladin Province, and 15 specimens from Al- Zubaydiyah sub-district, Wasit Province. The DNA sequences of C. kais were done using the mitochondrial DNA cytochrome b (cytb) gene. After analysis, the sequences were compared with sequences of ot
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Electro-kinetic remediation technology is one of the developing technologies that offer great promise for the cleanup of soils contaminated with heavy metals. A numerical model was formulated to simulate copper (Cu) transport under an electric field using one-dimensional diffusion-advection equations describing the contaminant transport driven by chemical and electrical gradients in soil during the electro-kinetic remediation as a function of time and space. This model included complex physicochemical factors affecting the transport phenomena, such as soil pH value, aqueous phase reaction, adsorption, and precipitation. One-dimensional finitedifference computer program successfully predicted meaningful values for soil pH profiles and Cu
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Objective(s) : This study aimed at evaluating the seroprevalence of anti -HCV and studying the
correlation between hemophilia and risk factors for acquiring HCV such as age , marital status &
occupation among hemophilic patients .
Methodology : 210 hemophilic patients in children welfare teaching hospital/medical city/Baghdad–Iraq
(hemophilia center) were investigated using prepared questionnaire and tested for HCV infection, those
were measuring patient’s age, hemophilia types and severity, marital status, residency and history of
previous HCV infection .
Results : Most hemophilic patients were hemophilia A at severe , hemophilia was at age group 20 – 29
years , the majority of patients were unmarried a
Ultrasonic extraction is an inexpensive, simple and efficient alternative to conventional extraction techniques, as compared with other novel extraction techniques such as microwave-assisted extraction & supercritical fluid extraction techniques, the ultrasound apparatus is cheaper and its operation is easier. Ultrasound assisted extraction has risen rapidly in the latest decade, and for most applications it has proven to be effective compared to traditional extraction techniques. In this paper, a method of ultrasonic-assisted extraction was used to extract Inulin from tubers of Jerusalem artichoke, which have been reported to have several medicinal properties and uses. Inulin is a storage carbohydrate found in many plants especially
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