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.
This paper proposes an on-line adaptive digital Proportional Integral Derivative (PID) control algorithm based on Field Programmable Gate Array (FPGA) for Proton Exchange Membrane Fuel Cell (PEMFC) Model. This research aims to design and implement Neural Network like a digital PID using FPGA in order to generate the best value of the hydrogen partial pressure action (PH2) to control the stack terminal output voltage of the (PEMFC) model during a variable load current applied. The on-line Particle Swarm Optimization (PSO) algorithm is used for finding and tuning the optimal value of the digital PID-NN controller (kp, ki, and kd) parameters that improve the dynamic behavior of the closed-loop digital control fue
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The aim of the present study was to isolated the Enterococcus spp. from milk samples of cow and vaginal swabs from aborted women and patient women in Baghdad during September 2016 to april 2017. All 100 milk sample collecting was carried out on California Mastitis Test (CMT) and the positive Percentage of CMT reactions was 5% and the percentage of Enterococcus isolates from mastitic milk was 60% and 30% from nonmasitic milk. The prevalence of Enterococcus spp was 31% of milk samples and the prevalence of Enterococcus spp. Isolates were 67.74% of the isolates of cow milk samples were Enterococcus faecalis, 25.80% was Group D and 6.45% was non groupable while Enterococcus spp. isolates from aborted women samples were 20% and all isolated was
... Show MoreA many risk challenge in (settings hospital) are multi- bacteria are antibiotic-resistant. Some type strains that ability adhesion surface-attached bio-film census. Fifteen MRSA isolates were considered as high biofilm producers Moreover all MRSA isolates; M3, M5, M7 and M11 produced biofilms but the thickest biofilm seen M7strain. The MIC values of N. sativa oil against clinical isolates of MRSA were between (0.25, 0.5, 0.75, 1.0) μg/ml While MRSAcin (50, 75, 100, 125) µg\ ml. All biofilms treated with MRSAcin and Nigella sativa developed a presence of live cells after cultured on plate agar with inhibition zone between MIC (18 – 15) and (14- 11)mm respectively.Yet, results showed that MRSA supernatant developed a inhibitory ef
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A total of 533 specimens were collected in survey of Brachyceran species from different regions of Iraq during February to November 2014 .This study was reported 16 species belonging to 13 genera and 7 families, the results showed that Dicranosepsis Duda, 1926 (Family; Sepsidae) is recorded the genus for the first time in Iraq.
The measurement of minority carrier lifetime (MCLT) ofp-n Si fabricated with aid of laser doping technique was reported. The measurement is achieved by using open circuit voltage decay (OCVD) technique. The experiment data confirms that the value of MCLT and proftle of Voc decay were very sensitive to the doping laser energy.
Surveying 59 references concerning the occurrence of the monogeneans of the family Diplozoidae parasitizing fishes of Iraq showed the occurrence of 15 valid species of this family which included one species of Diplozoon, one species of Eudiplozoon and 13 species of Paradiplozoon. In addition to these species, some unidentified adult and larval (diporpa larvae) specimens of the genus Diplozoon were reported from 12 fish hosts among which four fish species showed no infection with any of the nominated diplozoid species while the others showed mixed diplozoid infections. These diplozoids were reported from 27 fish host species in Iraq. All the diplozoids were recorded from freshwater habitats except one <
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Abstract: Recombinant Newcastle disease virus (rNDV) has shown an anticancer effect in preclinical studies, but has never been tested in a lung cancer models. In this study we explored the anticancer activity of genetically modified NDV expressing IL-2-P53 (rClone30–IL-2-P53) in lung cancer model. We have cloned IL-2 and P53 genes and inserted them in the viral genome of New Castle Disease Virus to create a genetically modified rNDV- IL-2-P53 virus and tested the anti-tumor activity of the new virus in vitro on different types of cancer cell lines by MTT assay. TheIL-2 and P53 gene were successfully cloned and inserted into the viral genome by using a Mlu I and Sfi I endonucleases, viral vector was constructed correctly and successf
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The current research is attempt to test the reflection of the lean management on the human resources management practices of two of the most important communication companies operating in Iraq (`Zain & Asia cell), The research aims to Determine the extent of adoption of the lean management approach in the two researched companies, as it improving human resource management practices. The research problem represented in the existence of lack of in some aspects of the application the lean management approach in service sector and neglecting the impact of its tools on the human resource management practices. For this purpose three principle research hypotheses has been formulated, first there is a correlation rel
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