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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BACKGROUND: Sickle cell nephropathy, a heterogeneous group of renal abnormalities resulting from complex interactions of sickle cell disease (SCD)-related factors and non-SCD phenotype characteristics, is associated with an increased risk for morbidity and mortality. AIMS: The aims of this study were to determine the frequency of microalbuminuria (MA) among pediatric patients with SCD and to determine risk factors for MA among those patients. SUBJECTS AND METHODS: A case–control study was carried out on 120 patients with SCD, 2–18 years old, registered at Basrah Center for Hereditary Blood Diseases, and 132 age-and sex-matched healthy children were included as a control group. Investigations included complete blood panel, blood urea, se
... Show MoreBackground: There are various secreted proteins affecting the prognosis of oral squamous cell carcinoma (OSCC) and one of them is Angiopoietin-2(Ang-2) which is thought to have an essential role in the development and progression of the tumor. Aim of the study: This study was conducted to determine the expression of (Ang-2) in (OSCC) to assess its correlations with clinicopathological parameters of the tumor. Material and Methods: 36 formalin- fixed, paraffin- embedded tissue blocks histologically diagnosed as OSCC were examined for Ang-2 immunohistochemical expression semi quantitively. Results: The expression of Ang-2 was significantly associated with histopathological grade (P value=0.023), while there is no significant association wi
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High cost of qualifying library standard cells on silicon wafer limits the number of test circuits on the test chip. This paper proposes a technique to share common load circuits among test circuits to reduce the silicon area. By enabling the load sharing, number of transistors for the common load can be reduced significantly. Results show up to 80% reduction in silicon area due to load area reduction.
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This study was conducted on five kinds of local soybean seeds (Ibaa, Hawija, Taqa.2, Lee74 and Hassan). The chemical analysis results showed that Hawija soybean has the highest percent of protein which was 38-08%, The amino acid percent was also higher than the other kinds(lysine, Thereonine and Tryptopham), and being 389,250,81 mg/gm nitrogen respectively Both amino acids were important for child nutrition. Hawija was selected, being the best for proteins and basic amino acids, and was utilized in preparation of the adjunct baby food formula. Eighteen formulas had been prepared by using soybean flour kind(Hawija), wheat flour kind (Abu gharib) and full fat powder milk (NIDO). Each formula contained 20% protein as recommended by F.A.O, W.
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The research aims to demonstrate the quality of the auditor’s report by analyzing a number of models represented by the auditor’s report based on the Iraqi audit evidence and the auditor’s report in light of the application of international auditing standards for the report and the auditor’s report in light of the application of the International Auditing Standard 701, in addition to explaining the impact of applying the International Auditing Standard 701. International Auditing 701 in enhancing the quality of the auditor's report, and in order to reach these goals, a comparison was made on international experiences before and after the application of the standard in addition to the results of the questionnaire distribut
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This study was conducted to investigate the relationship between corticosterone hormone in the blood of local Iraqi white chicken mothers and the sexual ratio of the hatching chicks. A field experiment located at the Poultry Research Station, Livestock Department, Agricultural Research Department, Ministry of Agriculture for the period from 16 of September 2018 to 25 of December 2018. One hundred and forty birds of Iraqi local chicken mothers (100 females+ 40 males) at the age of 28 weeks were provided from the poultry research station. The birds were raised in individual cages and distributed sequentially to the cages after the numbering of the females. Data were recorded in three periods, each period was of 28 days, and then the general a
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