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.
Keys for 22 species representing 10 genera of Thripidae were provided collection of
samples carried out during 1999-2001 in different localities in the middle of Iraq. Of them
four species are described as new to science, Frankliniella megacephala sp. nov; Retithrips
bagdadensis sp. nov; Chirothrips imperatus sp. nov; Taeniothrips tigridis sp. nov; Another
fourteen species are recorded for the first time in Iraq; Thrips meridionalis (Pri.);
Microcephalothrips abdominils (Crawford Scolothrips sexmaculatus (Pergande),);Scolothrips
pallidus (Beach); Scritothrips mangiferae Pri.; Frankliniella tritici Bagnall; Frankliniella
schultzie Trybom; Frankliniella unicolor Morgan; Retithrips aegypticus Marchal; Retithrips
java
In this paper, two types of iron oxide nanomaterial (Fe3O4) and nanocomposite (T-Fe3O4) were created from the bio-waste mass of tangerine peel. These two materials were utilized for adsorption tests to remove cefixime (CFX) from an aqueous solution. Before the adsorption application, both adsorbents have been characterized by various characterizations such as XRD, FTIR, VSM, TEM, and FESEM. The mesoporous nano-crystalline structure of Fe3O4 and T-Fe3O4 nanocomposite with less than 100-nm diameter is confirmed. The adsorption of the obtained adsorbents was evaluated for CFX removal by adjusting several operation parameters to optimize the removal. The optimal conditions for CFX removal were found to be an initial concentration of 40 and 50 m
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FLI1 is a member of ETS family of transcription factors that regulate a variety of normal biologic activities including cell proliferation, differentiation, and apoptosis. The expression of FLI1 and its correlation with well-known breast cancer prognostic markers (ER, PR and HER2) was determined in primary breast tumors as well as four breast cancer lines including: MCF-7, T47D, MDA-MB-231 and MDA-MB-468 using RT-qPCR with either 18S rRNA or ACTB (β-actin) for normalization of data. FLI1 mRNA level was decreased in the breast cancer cell lines under study compared to the normal breast tissue; however, Jurkat cells, which were used as a positive control, showed overexpression compared to the normal breast. Regarding primary breast carcinoma
... Show MoreBackground: Invasion in oral cancer involves alterations in cell-cell and cell-matrix interactions that accompanied by loss of cell adhesion. Catenins stabilize cellular adherence junctions by binding to E-cadherin, which further mediates cell-cell adhesion and regulates proliferation and differentiation of epithelial cells. The Wnt/β-catenin pathway is one of the major signaling pathways in cell proliferation, oncogenesis, and epithelial-mesenchymal transition. Aims of the study: to detect immunohistochemical distribution pattern and different subcellular localization of β-catenin in oral squamous cell carcinoma and relate such expression to Bryne’s invasive grading system. Materials and Methods: This study included 30 paraffi
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(1)
The current study is concerned over the benefits from our Prophet ' Surah , these benefits are seven which are : the first : ( The losing , disappointment , weakness in the affairs of those disbelievers ,the second : showing the affairs of the believers in respect to integrity , forgiveness and rest, the third : Indicating the big difference between the believers and disbelievers , each team has his own characteristics that being distinguished from the other, I have explained the benefit that the disbelievers are in a difficult
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Flavonoids were extracted from Zizyphus spina-christi leaves by Ethyl acetate after acid digested and used as antioxidant. The dried extract was added separately to each sample of fat extracted from hallow cow and sheep bones as follows: T1 cow fat, T2 control for cow fat, T3 sheep fat and T4 control for sheep fat (the control T2 and T4 reffered to samples without added antioxidant).
Samples were stored at -18, 5, 25 and 55 °C for 28 days. The storage trials were conducted at -18, 5 and 25 °C for 28 days for T1, T2, T3 and T4. The chemical indices examined initially and at the end of storage period. PVs was 1.46, 1.46, 1.8 and 1.8 meq/ Kg oil respectively, FFA values were 0.245, 0.245, 0.244 and 0.244% respectively and TBA va
In this study, polymeric composites were prepared from unsaturated polyester as a base material with glass powder (fluorescent) in different weight ratios (4, 6, 8, 10,and 11%) as a support material and after comparison before and after reinforcement of the prepared composites, an increase was found. In the values of mechanical properties (hardness, compressive strength), the shock resistance values decreased, but an increase in temperature leads to an increase in the values of shock resistance, as well as the values of compressive strength And it reduces the hardness value.
Results showed that the optimum conditions for production of inulunase from isolate Kluyveromyces marxianus AY2 by submerged culture could be achieved by using inulin as carbon source at a concentration of 2% with mixture of yeast extract and ammonium sulphate in a ratio of 1:1 in a concentration of 1% at initial pH 5.5 after incubation for 42 hours at 30ºC.
Concentrations of heavy metals (Copper Cu, Iron Fe, Manganese Mn, Cadmium Cd, and Lead Pb) have been studied in river crab Sesarma boulengeri (Outer part of the shield and interior tissues) which caught from two stations in Shatt Al – Arab river (Salhia and Aldeir areas). Elements concentrations were measured by Flame Atomic Absorption Spectrophotometer, concentration of heavy metals in the internal tissues was higher than in the outer shield in both of the stations with the highest value of the elements was to iron 95.21 mg\ kg during the spring as well as copper was 55 mg\kg and manganese was 39.09 mg\kg. The study showed the presence of seasonal changes in the studied heavy metals concentrations values in the tissues of river crab;
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