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Purification of G-Protein Coupled Receptor from Whole Cell of Local Strain of Saccharomyces cerevisiae
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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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Publication Date
Thu Dec 01 2016
Journal Name
2016 Asia-pacific Microwave Conference (apmc)
High selectivity microstrip bandpass filter using feedback CRLH-TL unit cell
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Publication Date
Mon Dec 24 2018
Journal Name
Nano Letters
Personalized Nanotherapy by Specifically Targeting Cell Organelles To Improve Vascular Hypertension
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Publication Date
Thu Jul 01 2021
Journal Name
Turkish Journal Of Physiotherapy And Rehabilitation
ORGANIZATIONAL CONFIDENCE AND ITS IMPACT ON THE DECENTRALIZED PERFORMANCE OF THOSE IN CHARGE OF MANAGING LOCAL CHAMPIONSHIPS FOR THE IRAQI ATHLETICS FEDERATION
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The aim of the research is to identify the level of both organizational trust and decentralized performance of those in charge of managing local championships for the Iraqi Athletics Federation, and to identify the effect of organizational trust in decentralized performance from their point of view. The descriptive approach was based on the method of relational relations on a sample of those in charge of managing local championships for the Iraqi Federation In athletics, represented by each of (coaches, referees, members, president and members of the administrative body of the central federation, and the president of members of sub-federations) for the sports season (2020/2021) of 260 individuals, all of them were intentionally chosen by (1

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Publication Date
Sat Jan 01 2022
Journal Name
Materials Today: Proceedings
Comparative study between the individual, dual and triple addition of (SF), (TGP) and (PVA) for improving Local Plaster of Paris (LPOP) properties
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Publication Date
Mon Jun 02 2008
Journal Name
Iraqi Postgraduate Medical Journal
[PDF] from iasj.net Isolation of tRNA From Uropathogenic Escherichia Coli
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Thirty uropathogenic E. coli isolates were isolated from hospitalized and non hospitalized patients, complaining of urinary tract infections, of Al-Kadhymia Teaching Hospital and subjected to tRNA extraction. A method of tRNA extraction was modified by adding sodium dodecyl sulfate (SDS) instead of urea. Polyacrylamide gel electrophoresis and two methods of staining, ethidium bromide staining and silver staining, as well as spectrophotometric detection were used.

Publication Date
Thu Oct 01 2020
Journal Name
August
INFLUENCE OF SOME FACTOR ON SOMATIC EMBRYOS INDUCTION AND GERMINATION OF DATE PALM BARHI C.V BY USING CELL SUSPENSION CULTURE TECHNIQUE
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INFLUENCE OF SOME FACTOR ON SOMATIC EMBRYOS INDUCTION AND GERMINATION OF DATE PALM BARHI C.V BY USING CELL SUSPENSION CULTURE TECHNIQUE

Publication Date
Fri Mar 27 2020
Journal Name
Plant Archives 20 (supplement 1), 1666-1670‏
INFLUENCE OF SOME FACTOR ON SOMATIC EMBRYOS INDUCTION AND GERMINATION OF DATE PALM CV BARHI BY USING CELL SUSPENSION CULTURE TECHNIQUEث‏
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Publication Date
Fri Mar 20 2020
Journal Name
Plant Archives (09725210) 20 (2)‏
Influence of some factor on somatic embryos induction and germination of date palm CV barhi by using cell suspension culture technique.‏
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Publication Date
Thu Sep 13 2018
Journal Name
Baghdad Science Journal
The Prognostic Value of some Epithelial-Mesenchymal Transition Markers and Metastasis-Related Markers in Human Transitional Cell Carcinoma of the Bladder
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Recent reports provided evidence that epithelial to mesenchymal transition (EMT) and some matrix metalloproteinases (MMPs) contribute to the invasion and metastasis of cancer cells. This study investigated the expression pattern of some EMT markers (E-cadherin and Vimentin) and some MMPs (MMP-2 and MMP-9) in transitional cell carcinoma (TCC). Fifty five paraffin embedded biopsies were included in this study. Expression pattern of E-cadherin and Vimentin was evaluated by immunohistochemistry while cytoplasmic mRNA expression of both MMP-2 and MMP-9 were determined by in situ hybridization. The expression of all markers were significantly increased with the increase of patient's age (? 50 years), and furthermore an increase in men expression

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Publication Date
Sun Jan 01 2023
Journal Name
Plant Archives
INFLUENCE OF SOME FACTOR ON SOMATIC EMBRYOS INDUCTION AND GERMINATION OF DATE PALM CV BARHI BY USING CELL SUSPENSION CULTURE TECHNIQUEe
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INFLUENCE OF SOME FACTOR ON SOMATIC EMBRYOS INDUCTION AND GERMINATION OF DATE PALM CV BARHI BY USING CELL SUSPENSION CULTURE TECHNIQUEe