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.
The present work aimed to investigate the neuraminidase (nan1) gene expression in 32 different clinical isolates of Pseudomonas aeruginosa to explore the role of the enzyme in different types of infection and might give a better understanding of host cell-pathogens interaction. In addition, the effect of monosaccharide D-mannose on neuraminidase gene expression in eight isolates was studied by utilizing a reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The results demonstrated that the highest expression of nan1 gene was in otitis samples (208,913.81) which were significantly higher than that from other infections (P < 0.01). While, the concentrations of gene copies obtained from urin
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Abstract
This study aims to evaluated the user satisfaction of retrieval services
concerning to universities thesis and dissertations in university dissertation
unit of Baghdad Library for achieving the following objectives:
1- Evaluating the performance of this unit (thesis unit of Baghdad University
Library) regarding to users opinion.
2- Recognizing the reasons in this unit behind the case of non satisfaction of
its users and trying to find the suitable solutions.
To achieve those two objectives, the questionnaire tool was performed
and determined the user's satisfaction level by using a sample survey. 1118
graduated students were subjected to this experiment. The following main
results were appeared:<
The objective of present study was to investigate the effect of using mixture volaticle oil of rosmarinus and nigella sativa to improve some of the meat quality characteristics, physical and limited storage time of minced cold poultry meat. Duplex volaticle oil was added at 0.025, 0.050 and 0.075 g/kg to minced poultry meat, these treatments were stored individually for 0 , 4 and 7 days at 4-7C0. After making several chemical, physical and oxidation indicators, the following results were obtained:
The process of adding volaticle oil to minced poultry meat led to significant increase (P<0.01)in moisture, prot
... Show MoreActivity test of the inhibitors purified from barley and broad beans crop proved the inhibition activity against 6 types of rots Pencillium ssp and Aspergellusflavus and Aspergillus niger and Fusarium solani and Fusarium semitectum and Mucor with three concentrations 0.1 and 0.2 and 0.3 mg/ml, where the inhibitor purified from the second peak of broad beans proved that it had a higher inhibition activity against the growth of test rots which were 53.75 and 62.5 and 78.5 and 76.25 and 84 and 18.8% respectively, at 0.3 mg/ ml followed by the first peak of the inhibitor purified from broad beans the inhibition activity were 43.75 and 50 and 62.96 and 75 and 80 and 12.5 then the inhibitor purified from barley in which the inhibition activity
... Show MoreThe surplus glycerol produced from biodiesel production process as a by-product with high quantity can be considered as a good source to prepare glycerol carbonate (GC) whereas with each 1000 kg from biodiesel obtains 100 kg from glycerol. Glycerol converted to glycerol carbonate over bio-char as a catalyst prepared by slow pyrolysis process under various temperatures from 400 ᴼC to 800 ᴼC. The char prepared at 700 ᴼC considered as a best one between the others which was manufactured to activate the transesterification reaction. GC have large scale of uses such as liquid membrane in gas separation, surfactants ,detergents , blowing agent , in plastics industry, in Pharmaceutical industry and electrolytes in lithium batteries.
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This research involved synthesis of new β-Lactam derivative from Azo compound[4-amino-N-(pyrimidine-2-yl)-3-(pyrimidine-2-yldiazenyl) benzene sulfonamide] (S1) record previously by many steps. Starting conversion the free amino group in an azo comp. to chloro acetamide derivative(S2), then reacted it with urea to give the oxazole ring derivative (S3) that which containing free amino group. The condensation reaction between the amino group and P-bromobenzaldehyde to produce Shiff base (B14). Finally staudinger's cyclo addition reaction go run between the Shiff base derivative (B14) and chloro acetyl chloride in the presence of tri ethyl amine (Et3N) as Base catalyst and dioxane a
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This research includes study of the effect of two kinds of Anthocyanin extracted , from extracted orange fruit ( Anthocyanin Evolvulus ,Methiola Violet ) on two types of pathological bacteria E.coli , staphylococcus aureus. The result shows that two kinds of extraction have nearly similar effect , and there is Inhibition zone of no growth between 10-12mm ,and the extraction (1) that has concentration of 10-3 mol./L is more effective..
Keys for 22 species representing ten genera Thripidae collection carried out during 1999-2001 in different localities in the middle of Iraq. Of them four species are described as new to science, Frankliniella megacephala sp. nov; Retithrips bagdadensis sp. nov; Chirothrips imperatus sp. nov; Taeniothrips tigridis sp. nov; Another thirteen species are recorded for the first time in Iraq; Thrips meridionalis (Pri.); Microcephalothrips abdominils (Crawford); Scolothrips pallidus (Beach); Scritothrips mangiferae Pri.; Frankliniella tritici Bagnall; Frankliniella schultzie Trybom; Frankliniella unicolor Morgan; Retithrips aegypticus Mar
... Show MoreThe emergence of staphylococci, either coagulase negative (CNS) or coagulase positive (CPS), as important human pathogens has implied that reliable methods for their identification are of large significance in understanding the diseases caused by them. The identification and characterization of staphylococci from biopsies taken from human breast tumors is reported here. Out of 32 tissue biopsies, a total of 12 suspected staphylococci grew on mannitol salt agar (MSA) medium, including 7 fermenters and 5 non-fermenter staphylococci based on traditional laboratory methods. Polymerase chain reaction (PCR) successfully identified seven isolates at the genus level as methicillin resistant Staphylococcus spp. by targeting a common region of the me
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