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.
A study of the Torymid collection of Iraq. resulted in undescribed species of the genus
Liodontonierus Gah. L. longicorpus sp. n. with 2 figures.
(1)
(1)
Recently, important efforts have been made in an attempt to search for the cheapest and ecofriendly alternatives adsorbents. In the present work, waste molasses from Iraqi date palm (Zahdi) had been used as a provenance to produce charcoal for the removal of methylene blue (MB) dye from water. The optimum prepared charcoal was obtained at 150 C, by increasing temperature to 175 C, the charcoal had almost converted to ash. The obtained charcoal have been inspected for properties using scanning electron microscope (SEM), atomic force microscope (AFM), porosity and surface area. Adsorption data were optimized to Langmuir and Freundlich and adsorption parameters have been evaluated. The thermodynamic parameters like a change
... Show More
(1)
After the fall of the Third Dynasty of Ur at the hands of the Elamites during the reign of its last ruler, King Ibi-Sin, the Amorites continued to flow more into Mesopotamia in the form of large migrations that established a number of contemporary and conflicting ruling dynasties that formed the Old Babylonian era. Among these dynasties was the Larsa dynasty, founded by King Nablanum in the city of Larsa. The fourth king who ruled in the Larsa dynasty, Gungunum, was one of the most powerful kings who stood up to the Isin dynasty, which had until that time enjoyed great political superiority in the country. The kings who ruled after him expanded their influence and controlled a number of cities, especially the cities of Nippur and Uruk, whic
... Show MoreThis study aimed to extraction of essential oil from peppermint leaves by using hydro distillation methods. In the peppermint oil extraction with hydro distillation method is studied the effect of the extraction temperature to the yield of peppermint oil. Besides it also studied the kinetics during the extraction process. Then, 2nd -order mechanism was adopted in the model of hydro distillation for estimation many parameters such as the initial extraction rate, capacity of extraction and the constant rat of extraction with various temperature. The same model was also used to estimate the activation energy. The results showed a spontaneous process, since the Gibbs free energy had a value negative sign.
(7)
In this work ,medical zinc oxide was produced from zinc scraps instead of traditional method which used for medical applications such as skin diseases, Iraq is importing around 50 ton/year for samarra plant the producted powder has apartical size less than 5 micron and the purity was more than 99.98%,also apilot plant of yield capacitiy 15 kg/8hours wsa designed and manufactured .
Trickle irrigation is one of the most conservative irrigation techniques since it implies supplying water directly on the soil through emitters. Emitters dissipate energy of water at the end of the trickle irrigation system and provide water at emission points. The area wetted by an emitter depends upon the discharge of emitter, soil texture, initial soil water content, and soil permeability. The objectives of this research were to predict water distribution profiles through different soils for different conditions and quantify the distribution profiles in terms of main characteristics of soil and emitter. The wetting patterns were simulated at the end of each hour for a total time of application of 12 hrs, emitter disch
... Show MoreA mixture of algae biomass (Chrysophyta, Cyanophyta, and Chlorophyte) has been investigated for its possible adsorption removal of cationic dyes (methylene blue, MB). Effect of pH (1-8), biosorbent dosage (0.2-2 g/100ml), agitated speed (100-300), particle size (1304-89μm), temperature (20-40˚C), initial dye concentration (20-300 mg/L), and sorption–desorption were investigated to assess the algal-dye sorption mechanism. Different pre-treatments, alkali, protonation, and CaCl2 have been experienced in order to enhance the adsorption capacity as well as the stability of the algal biomass. Equilibrium isotherm data were analyzed using Langmuir, Freundlich, and Temkin models. The maximum dye-sorption capacity was 26.65 mg/g at pH= 5, 25
... Show More
(19)