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.
SJ Mohammed, AA Noaimi, KE Sharquie, JM Karhoot, MS Jebur, JR Abood, A Al-Hamadani, Al-Qadisiyah Medical Journal, 2015 - Cited by 20
This study was aimed to one of the most prevalent causes for endodontic treatment failure is the presence of Enterococcus faecalis bacterium within teeth root canals. To achieve successful treatment, it is so important to study E. faecalis behavior. The aim of study was to investigate biofilm production and antibiotic sensitivity of E. faecalis isolated from root canals. Results showed isolation of E. feacalis (65%) of samples, identified by specific gene by PCR technique. Most isolates were sensitive to Imipenem and resistant to Erythromycin, Clindamycin, Tetracycline and Trimethoprim. Strong biofilm production was detected among 29.5% of highest antibiotic resistant isolates. The results may indicate that infected root canals with E. feac
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The degradation and mineralization of 4-chlorophenol (4-CP) by advanced oxidation processes (AOPs) was investigated in this work, using both of UV/H2O2 and photo-Fenton UV/H2O2/Fe+3 systems.The reaction was influenced by the input concentration of H2O2, the amount of the iron catalyst, the type of iron salt, the pH and the concentration of 4-CP. A colored solution of benzoquinon can be observed through the first 5 minutes of irradiation time for UV/H2O2 system when low concentration (0.01mol/L) of H2O2 was used. The colored solution of benzoquinon could also be observed through the first 5 minutes for the UV/H2O2/Fe+3 system at high
concentration (100ppm) of 4-CP. The results have shown that adding Fe+3 to the UV/H2O2 system enhanced
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Refractories are mineral and chemical-, based, materials with excellent heat resistance, making them ideal for use in the construction of ovens, furnace walls industries. According to this our research is concerned to study the effect of addition of (4% CaO) and (5% graphite) on the silica brick properties. Different amounts of CaO and Graphite were included in the white sand (raw ingredients) of silica bricks as a binder to prepare the composition then the composition were sintered using Different sintering temperatures ranging from (1000–1400)𝛐C under static air. Density, thermal conductivity, porosity, and water absorption Compression there was power tested after sintering. XRD analysis was used to identify raw materials’
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Through his female characters, Walt Disney was able to reflect the
development of the general outlook upon women in the Western world over a period
of sixty years.
This paper sheds light in how the cartoon characters changed in their reaction
to problems and means of solving them from the 30s through to the 90s of the
previous century. The main characters will be seen according to the films’ dates of
production and release.
Probiotics are live microbes that give many health benefits to human beings and animals, the most studied and commonly used probiotics are Gram-positive bacteria; lactobacilli and bifidobacteria. At nowadays, Lactobacillus spp. constitute more than two-thirds of the total numbers of probiotic species. The present study aimed to characterize Lactobacillus that locally isolated from human mouth and feces as probiotics. A total of three Lactobacillus isolates; Lactobacillus fermentum Lb2, Lactobacillus rhamnosus Lb9, and Lactobacillus paracasei Lb10 were investigated in respect to acid and bile salts tolerance, antibiotics susceptibility, and cell surface hydrophobicity in vitro using bacterial adhesion to hydrocarbons method. In compa
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The present study examines the extraction of lead (Pb), cadmium (Cd) and nickel (Ni) from a contaminated soil by washing process. Ethylenediaminetetraacetic acid disodium salt (Na2EDTA) and hydrochloric acid (HCl) solution were used as extractants. Soil washing is one of the most suitable in-situ/ ex-situ remediation method in removing heavy metals. Soil was artificially contaminated with 500 mg/kg (Pb , Cd and Ni ). A set of batch experiments were carried out at different conditions of extractant concentration , contact time, pH and agitation speed. The results showed that the maximum removal efficiencies of (Cd, Pb and Ni ) were (97, 88 and 24 )&nbs
... Show MoreBased on collections made during March to September 2012. A totals of 58 species belong to 33 genera were identified from extra north to extra south of Erbil governorate, among them 30 species are registered as a new record to flora of Iraq. Most attention was paid to the most common and abundant lichens that present almost in most locations, which were Collema cristatum, Diploschistes scruposus, Lecanora dispersa, Lecanora murales, Pertusaria flavicunda, Placidium lacinulatum, Thelomma californicum and Verrucaria Maura.
The removal of Anit-Inflammatory drugs, namely; Acetaminophen (ACTP), from wastewater by bulk liquid membrane (BLM) process using Aliquat 336 (QCl) as a carrier was investigated. The effects of several parameters on the extraction efficiency were studied in this research, such as the initial feed phase concentration (10-50) ppm of ACTP, stripping phase (NaCl) concentration (0.3,0.5,0.7 M), temperature (30-50oC), the volume ratio of feed phase to membrane phase (200-400ml/80ml), agitation speed of the feed phase (75-125 rpm), membrane stirring speed (0, 100, 150 rpm), carrier concentration (1, 5, 9 wt%), the pH of feed (2, 4, 6, 8, 10), and solvent type (CCl4 and n-Heptane). The study shows that high ext
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