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.
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The purpose of this study was to determine the effect of lupine flour (L.f) and lupine protein concentrate (L.P.C) incorporation on chemical, nutritional and sensual qualities characteristics of biscuit (L.P.C) was prepared by isoelectric precipitation method. A standard recipe for biscuit preparation by wheat patent flour used as the control. Wheat flour in the control treatment was replaced with (L.f) and (L.P.C) at levels 10, 20 & 30%. Chemical composition of (L.f), (L.P.C) and biscuit treatments were studied. Results showed that protein contents were 35.35 & 75.80% for (L.F) and (L.P.C), respectively. While they amounted to 14.70, 16.16 & 18.61% for (L.f) incorporated biscuits and
... Show MoreThis studies p- CuO / n - Si hete-rojunction was deposited by high vacuum thermal evaporation of Copper subjected to thermal oxidation at 300 oC on silicon. Surface morphology properties of The optical properties concerning the transmission spectra were studies for prepared thin films. this structure have been studied. XRD anaylsis discover that the peak at (𝟏𝟏𝟏-) and (111) plane are take over for the crystal quality of the CuO films. The band gap of CuO films is found to be 1.54 eV. The average grain size of is measured from AFM analysis is around 14.70 nm. The responsivity photodetector after deposited CuO appear increasing in response
The theory of general topology view for continuous mappings is general version and is applied for topological graph theory. Separation axioms can be regard as tools for distinguishing objects in information systems. Rough theory is one of map the topology to uncertainty. The aim of this work is to presented graph, continuity, separation properties and rough set to put a new approaches for uncertainty. For the introduce of various levels of approximations, we introduce several levels of continuity and separation axioms on graphs in Gm-closure approximation spaces.
Optoelectronic devices, widely used in high energy and nuclear physics applications, suffer severe radiation damage that leads to degradations in its efficiency. In this paper, the influence of gamma radiation (137Ce source) and beta radiation (90Sr source) on the photoelectric parameters of the Si solar cell, based on the I–V characterization at different irradiation exposer, has been studied. The penetrating radiation produces defects in the base material, may be activated during its lifetime, becoming traps for electron–hole pairs produced optically and, this will, decrease the efficiency of the solar cell. The main objective of the paper is to study and measure changes in the I–V characteristics of solar cells, such as efficienc
... Show MoreBackground: For various reasons, inguinal hernia repair under local anaesthesia is not well accepted to both patients and surgeons. The patients fear from pain and surgeons need full relaxation and co-operation to do successful hernia repairMethods: purpose of this study is to evaluate the effectiveness of local anaesthesia in inguinal hernia repair.prospective study was made from January 2011-0ctober 2013 , on a total of 50 patients with inguinal hernia operated on under local anaesthesia. Patients were selected primarily on the basis of their willingness to accept the procedure after the technique was described to them.Results: In this study 50 patient and 58 herniorrhaphies done for them during a period of about 34months were evaluate
... Show MoreThis study was conducted to investigate the presence of Staphylococcus aureus in the red and white meat available in local markets. They were selected ten samples of red and white meat randomly (Iraq, Saudi Arabia, Turkey, and Brazil) from different markets in Baghdad, and the results of reading the nutrition facts of media indication card showed that all models confirm to the Iraqi standard quality in terms of scanning all data of the media indication card, except for the birds of Bayader, where the date of expire & production date of the product was not mentioned. Also, the results of the study showed that there is no Staphylococcus aureus in local red and white meat as well as imported.
The development of better tools for diagnosis and more accurate prognosis of cancer includes the search for biomarkers; molecules whose presence, absence or change in quantity or structure is associated with a particular tumour or prognosis/therapeutic outcome. While biomarkers need not be functionally relevant, if cell survival, then they could also provide new targets for therapeutic drugs. In recent years attention has been applied to a group of proteins known as cancer testis antigens (CT antigens) [1]. These proteins are products of genes whose expression was normally confined to the testis, yet they are expressed in tumour cells. CT genes are bound to serve a wide array of roles in the testes, which have many highly differentiated cel
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