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.
Time is very important in educational institutions. It is also one of our contemporary problem ‚as time is a clear – cut and limited factor‚ it demands that administrators should monitor it by administering and monitoring the principles of time.
Hence‚ the researcher attempts to identify the skills of administrating time and the reasons that cause the waste of time of the Heads of Departments at university of Baghdad.
Significance of the research:
Time is very important to all educational administrators and one of them is the institutions of Higher education. One of the
... Show MoreIn this study the Bauxite has been activated and used to prepare two complexes: Bauxite - urea and Bauxite - melamine, these complexes were merged and polymerized with formaldehyde to prepare the complex Bauxite polymer - urea - melamine - formaldehyde (modified Bauxite). In the Bauxite-urea complex XRD results indicate that the urea molecules penetrate among the layers of the crystal plane (110) of the Gibbsite mineral while in the Bauxite-melamine the interaction was at the outer surface of the Bauxite forming minerals because the relatively large volume of the melamine molecule. FT-IR results show the interaction of these two bases with Bauxite was mainly based on the hydrogen bonding and in less extent on the coordination between N l
... Show MoreIn the present work , the a2 - ratio method has been used to calculate the multipole mixing ratios , 5 - values • of y - transitions from excited levels of deformation nucleiL-. ( 152Sm ) .
The results obtained confirm the validity of this method in calculating the o - values .
The present results are in good agreement with those of the experimental results, ref.( I ,2) , and of theoretical results using interaction boson model (IBM-I) ,ref. (5).
The personality of the hero Salah al-Din al-Ayyubi (may God have mercy on him) came from the womb of jihad after difficult travails that the Arab Islamic nation experienced through the jihad of its loyal and honest sons who vowed themselves to God in defense of his religion and law, so between 490 AH - 540 AH outstanding jihadi leaders emerged who took upon themselves the responsibility of jihad and mobilizing the nation's energies To fight its enemies - the Franks, the Crusaders - in the Levant, and those leaders succeeded in achieving impressive victories over the Frankish military effort and regained some cities that were usurped by the Franks. Balak bin Bahram, Suqman, and Jakarmish, but these leaders could not maintain a state of un
... Show MoreAnaerobic digestion is a technology widely used for treatment of organic waste for biogas production as a source for clean energy. In this study, poultry house wastes (PHW) material was examined as a source for biogas production. The effects of inoculum addition, pretreatment of the substrate, and temperature on the biogas production were taken into full consideration. Results revealed that the effect of inoculum addition was more significant than the alkaline pretreatment of raw waste materials. The biogas recovery from inoculated waste materials exceeds its production from wastes without inoculation by approximately 70% at mesophilic conditions. Whereby, the increase of biogas recovery from pretreated wastes was by 20% higher than its
... Show More
Background: Determination of local bone mineral density (BMD) immediately after implant insertion play an important role in implant success rate, may offer comprehensive description of the bone, and give enough information to the surgeon prior to implant insertion and at follow up status. The aim of the present study is to evaluate the changes of local bone density in the dental implant recipient sites by using computerized tomography. Material and method: The sample consisted of (20) dental implants recipient sites, bone density assessment was done twice, immediately after implants insertion and after six months. Results: The mean HU of the bone around the implant insertion site, immediately after implant placement was 552.28 HU, and inc
... Show MoreThis article conclude a theoretical study for the possibility to produce additional electric power from Iraqi steam power plants by cutting – off high-pressure feed water heaters . Three separated steam power plants which Dura , south –Baghdad and Nasria were studied . The investigation showed the possibity of increasing the electric power from 10 to 15% for Dura and Nasria , whereas 6% for south – Baghdad . According to the nowadays of operation to Iraqi steam power plants , the results showed that by cutting–off high pressure feed water heaters we can generate additional electric power about 250 MW during 3-4 hrs. daily. In addition, the fuel consumption can be reduced in comparison with diesel generat
... Show MoreThe δ-mixing ratios have been calculated for several γ-transitions in 90Mo using the 𝛔 𝐉 method. The results are compared with other references the agreement is found to be very good .this confirms the validity of the 𝛔 𝐉 method as a tool for analyzing the angular distribution of γ-ray. Key word: population parameter, γ-ray transition, 𝛔 𝐉 method, multiple mixing ratios.