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.
A total of 200 samples (180 fecal materials and 20 organ samples) were collected from (5 different poultry farms, 10 local poultry shops, 5 houses poultry, 5 Eggs stores shops and 5hand slaughters centers) in Ibb city, Yemen, 2014. According to morphological, cultural, as well as biochemical characterization and serological tests, 59(29.5%) isolates were identified as Salmonella spp. and all Salmonella isolates were categorized by serotype, which comprised of, 37(62.71%) Salmonella Typhimurium serovar, 21(35.59%). Salmonella Enteritidis serovar and 1(1.69%) Salmonella Heidlberg serovar. Antibiotic sensitivity test was done for bacterial isolates and the results showed there were clear differences in antibiotic resistant. Antimicrobial
... Show MorePseudomonas aeruginosa readily binds to different kind of abiotic surfaces and form biofilm. The ability of the bacterial species to form biofilm onto polyvinyl chloride (PVC) is associated with several economic, health and environmental problems. The effect of kind of water on ability of this bacterium to form biofilm is scanty in literature. In present study, the ability of different environmental isolates of P. aeruginosa to form biofilm onto polystyrene microtiter plate was evaluated. Furthermore, the effect of waters that collected from different sources on biofilm formation of this bacterium onto PVC was studied. Spectrophotometric method was used to check the ability of bacteria to form biofilm and evaluated the role of waters onto a
... Show MoreThe quantum chromodynamics theory approach was taken to study the photonic emission from interaction of quark gluon at high at Bremsstrahlung processes. Strength coupling, quark charge 𝑒𝑞 , flavor number 𝑛𝐹 , thermal energy T of system, fugacity of gluon ƛ𝑔, fugacity of quark ƛ𝑞 , critical temperature 𝑇𝐶 and photons energy 𝐸 are taken to calculate photons rate via the quantum system. Photons emission rate studies and calculates via high energy 400MeV to 650 MeV using flavor number 3 and 7 for 𝑢̅𝑔 → 𝑑̅𝑔𝛾 and 𝑐𝑔 → 𝑠𝑔𝛾 systems at bremsstrahlung processes with critical temperature (𝑇𝑐 = 190 and 196) MeV with photons energy (1-10) GeV. The confinement and de-confineme
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The study area comprises Injana Formation (Late Miocene), exposed on the hills nearby of Tharthar Lake and about 120 km north of Baghdad city. This study depends on sedimentologic and facies analysis to recognize paleoenvironment and recognize the kinds of vertebrate bone fossils during Late Miocene. Sedimentologic and facies analysis showed many sedimentary facies: facies (Se) of scoured erosional surface, facies of (Sp) cross- bedded sandstones, facies (Fs) of fine sandstone facies, facies of (Fc) claystone, and facies of (C) calcareous clay. Facies analysis referred to the sub environments which are: point bar, over bank and floodplain in addition to fining upward cycles of deposition, which refers to meandering flu
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In Present study, 25 clinical isolates of Proteus spp. of clinical samples, urine, wounds and burns collected from different hospitals in Baghdad city, all isolates were identified as Proteus mirabilis using different bacteriological media, biochemical assays and Vitek-2 system. It was found that 15 (60%) isolates were identifying as P. mirabilis. The susceptibility of P. mirabilis isolates to cefotaxime was 66.6 %, while to ceftazidime was 20%. Extended spectrum β-lactamses producing Proteus was 30.7 %. DNA of 5 isolates of P. mirabilis was extracted and detection for blaVEB-1 gene by using multiplex polymerase chain reaction (PCR). Results showed that the presence of this gene in all tested isolates, as an important indicator for increas
... Show MoreThe study included isolate and diagnose fungus Fusarium solani of the local soil and purified and development in the PDB medium and the filtrate extracted using a solvent (Ethyl acetate) to obtain the fungal secondary metabolites extract. This extract has shown bioactivity against both reference isolates (E.coli (ATCC25922) and S.aureus(NCTC6571)) and pathogenic isolates S.pyogenes, K. pneumonia and S.typhimurium using agar disk diffusion technique , The diameters of the inhibition zones of fungal secondary metabolites24.0 mm against E.coli and 31.5 mm against S.aureus,and 34.0 mm against K.pneumoniae and 18.0 mm against S.pyogenes and 33.5mm against S.typhimurium. The test revealed the minimum inhibitory concentration (MIC) of the fungal
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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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Cyprinidae 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
Background: Salvia officinalis is a plant belong to
Labiatae family .The common name of Salvia is sage
which mean save. The leaves of Salvia have special
oil which is effective against filamentous fungi and
yeasts such as Candida albicans which is the
causative agent of vaginal candidiasis in women
Methods. Cultures from 50 swabs of Candida
albicans isolated from vagina of 70 patient women
who complains from vaginal problems, their ages
(24-43) years from Central City Hospital during
Febreoury 2009 to April 2009 were cultured on
Sabouraud Dextrose Agar (SDA) .Nystatin was used
as positive reference standard to determine the
sensitivity of this fungus . and less this concentration
there was no min