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.
(7)
Five different bacterial isolates [ Vibrio cholera (Ogawa) , Vibrio cholera (Inaba) , Salmonella typhi , Salmonella paratyphi and ? Salmonella typhimurium ] were obtained from the Central Health Laboratory . Both sensitivity tests (MIC , MBC and wells method ) against these bacteria were performed by using the aqueous of leaves extract of Marjoram plant. The results cleared that the values of MIC for Vibrio cholera serotypes Ogawa and Inaba were 100 mg/ml , while the value of MBC was 200 mg/ml. The value of the Inhibition zone at 100 mg /ml concentration for both Ogawa and Inaba were 13 mm and 9 mm respectively. Our results showed that the three types of Salmonella didn’t show any inhibition zone at 200 mg/ml .
This research examines aspects of Imam Al-Qurtubi's efforts in matters related to the topics of faith that he dealt with in his interpretation (the provisions of the provisions of the Qur’an); and it has been shown that Imam Al-Qurtubi in most of the doctrinal issues dealt with in his interpretation approximates the doctrine of the Sunnis of Ash'aris, Maturids and people of impact. He had responses to some of the teams, such as stereoscopic, karami, and Mu'tazilite, reinforcing the responses with transfer and mental evidence.
It was found that Imam Al-Qurtubi says that there is an increase in faith and a decrease in it, that the increase is through actions (so obedience increases and decreases in sin) and that the
An investigation was conducted effect of addition co- solvent on solvent extraction process for two types of a lubricating oil fraction (spindle) and (SAE-30) obtained from vacuum distillation unit of lube oil plant of Daura Refinery. In this study two types of co-solvents ( formamide and N-methyl, 2, pyrrolidone) were blended with furfural to extract aromatic hydrocarbons which are the undesirable materials in raw lubricating oil, in order to improve the viscosity index, viscosity and yield of produced lubricating oil. The studied operating condition are extraction temperature range from 70 to 110 °C for formamide and 80 to 120 °C for N-methyl, 2, pyrrolidone, solvent to oil ratio range from 1:1 to 2:1 (wt./wt.) for furfural with form
... Show More
Background: Globally, breast cancer is the second leading cause of death among women in Iraq. Several genetic and environmental factors are associated..
This research includes a detaile description of new species Rhyncomya irakensis sp. nov.
from Iraq.
Localities distribution, host plants and data of collection were recorded.