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 letters of the Al-Safa Brothers contain many tales, in which each story is a literary text that is independent from the other texts, but it constitutes a unified entity in its entirety, because these stories contain recurring structural elements that go into building each one of them. The stories - the subject of the analysis - were not keen on something that was keen on adhering to the Sufi vision and its apparent effect on the tales of the messages, starting with the announcement of the narration to the narrator and his attempt to persuade the recipient of the truth of what is being told and its realism, and made him believe it. The stories correspond to the formal construction of the ideal style defined by th
... Show MorePyrolysis of virgin polyethylene plastics was studied in order to produce hydrocarbon liquid fuel. The pyrolysis process carried out for low and high-density polyethylene plastics in open system batch reactor in temperature range of 370 to 450°C.
Thermo-gravimetric analysis of the virgin plastics showed that the degradation ranges were between 326 and 495 °C. The results showed that the optimum temperature range of pyrolysis of polyethylene plastics that gives highest liquid yield (with specific gravity between 0.7844 and 0.7865) was 390 to 410 °C with reaction time of about 35 minutes. Fourier Transform Infrared spectroscopy gave a quite evidence that the produced hydrocarbon liquid fuel consisted ma
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The development of a new, cheap, efficient, and ecofriendly adsorbents has become an important demand for the treatment of waste water, so nano silica is considered a good choice. A sample of nanosilica (NS) was prepared from sodium silicate as precursor and the nonionic surfactant Tween 20 as a template. The prepared sample was characterized using various characterization techniques such as FT-IR, AFM, SEM and EDX analysis. The spectrum of FTIR confirms the presence of silica in the sample, while SEM analysis of sample shows nanostructures with pore ranging (2-100nm).The adsorptive properties of this sample were studied by removing Congo red dye (CR) from aqueous solution. Batch experimental methods were carried o
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Six isolates of Bacillus thuringiensis were isolated from Iraqi soil characterized as non- insecticidal and non- hemolytic parasporal inclusion proteins. Bacterial isolates were propagated on nutrient broth. Then, the parasporal inclusion proteins were extracted and processed with proteinase K and trypsin. The major protein segments produced of 64KDa were characterized and tested for cytocidal activity against human leukemic T- cells (CLL) (Chronic lymphoid leukemia). Results indicated that the treated parasporal proteins of four isolates (Bt2, Bt3, Bt4 and Bt6) showed strong cytotoxicity with no significant differences between normal lymphocytes and leukemic lymphocytes. Two isolates BtA1 and BtA5 show discriminative cytotoxicity between n
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Soils at Al-Koot-Btera were choosen to determine their sedimentary
environments. It is found that there are , five soil series and as mentioned :
MF11-MW9-DM97-DM57-DF95 . The five found soil series are of internal
well drained ,moderate and imperfect. Their textures vary in moderately,fine
and moderately fine.
indicating that sediments rang from poorly to very poorly sorting. Values of median
diameter Md Ø range between 4.11-7.80 Ø .The relation between the sorting
and median diameter shows that 95.24% of samples is a sedimentary
environment of aquite river , while 4.76% is aeolian sediments.
The values of meso to platy kurtic of most horizon materials rang
between 0.67-1.26 Ø .That is to say the samples
In this research, Schiff bases derived from the reaction of anthrone with different heterocyclic amines have been described. The resulted Schiff base compounds were reacted with various nucleophiles in order to obtain new heterocyclic derivatives. Chemical structures of all products were confirmed by IR, 1H-, 13C-NMR spectral data and elemental analysis. All synthesized compounds were in vitro tested against a standard strain of pathogenic microorganism including Gram +ve bacteria (Staphylococcus aureus), Gram –ve bacteria (Escherichia coli), and fungi (Candida albicans).
Intestinal parasites present in freshwater from the Al- Fallujah, Al- Habbaniyah and Al-Alwarar, of the Euphrates river in Iraq are Cryptosporidium spp (25.3%), Giardia sp (3.3%), Eimeria sp (3.3%), Pinworm eggs (3.3%), Naegleria sp (15.3%), Lecane niwati (1.3%), Trichomonas hominis (19.3%), Acanthamoeba spp (24.6%), Entamoeba coli (20.6%), Balantidium coli (12%), Ascaris sp (3.3%), Volvox sp (26%), Chilomastix mesnili (4%), Pelomyxa palustris (2.6%), Trinema enchelys (2.6%), Actinophrys Sol (7.3%), Amobea Vespertilio (9.3%), Rhabditea (5.3%), paramecium bursaria (9.3%), cyst of cestode (6%), Oocyst protozoa (16%), Euglena gracilis (10.6%).were isolated. The study's goal was to isolate some of the parasites that pollute the Euphrat
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Cloud computing is the new technological trend for future generations. It represents a new way to use IT resources more efficiently. Cloud computing is one of the most technological models for developing and exploiting infrastructure resources in the world. Under the cloud, the user no longer needs to look for major financing to purchase infrastructure equipment as companies, especially small and medium-sized ones, can get the equipment as a service, rather than buying it as a product. The idea of cloud computing dates back to the sixties of the last century, but this idea did not come into actual application until the beginning of the third millennium, at the hands of technology companies such as Apple, Hp, IBM, which had
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