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.
Assessment of Salivary Macrophage Inflammatory Protein-1 Alpha Level in Different Stages of Periodontitis, Riyam Muthanna Muhammed*, Hadeel Mazin Akram
The aims of this study are to explore the commercial artifacts in the following three kinds of vegetables oils, Nigella Sativa, Trigonella foenum-graecum Linn,and Zingiber officinale. These oils have been very popular medicinal plants which are commonly used in traditional medicine .These commercial oils have been compared with the extracts of these plants.
The physical properties of extracts and commercial oils of these plants have been stuied. We observed that the refractive index of the plants matches and non-significant, while specific gravity of Nigella Sativa has similar specific gravity in both extracts and commercial oil in contrast with Trigonella foenum Linn,and Zingiber officinale and we found significant difference (P<
This study was conducted to estimate some heavy metals cadmium, lead, nickel and iron in 15 samples of Iraqi honey with 3 replicates for each sample which were collected from apiaries near potential contamination areas in five Iraqi governorates, including Baghdad, Karbala, Babylon, Diyala and Salah al-Din. The atomic absorption technique was used to estimate the concentrations of heavy metals, the results showed that there were significant differences at (P≤0.05) between the concentrations of these elements in the honey samples, the highest concentrations of cadmium 0.123 mg/kg were recorded in Baghdad, near the petrochemical production complex, lead 4.657 mg/kg and nickel 0.023 mg/kg in Babylon near the power plant, iron was
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Imidacloprid is systemic insecticide (1-[(6-chloro-3-pyridinyl) methyl]-N-nitro-2-imidazolidinimine) and the world’s most widely used has significant efficacy against a broad variety of pests and a unique mode of action by using it spreader and irrigation. The persistence of this pesticide in the soil means that it causes environmental damage that must be cleaned up. In this study collected and identified the best bacteria isolate that breakdown imidacloprid from the Plant Protection Director in Baghdad, which has been using neonicotinoid pesticides for years in their own greenhouse for pest control. Using high-performance liquid chromatography HPLC to measuring the residual concentrations of imidacloprid in MSM media at a concentration o
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To explore the durability of some local species of wood to fungal deterioration among the
storage period, this research has conducted on three species Eufcalyptus cammaldulensis,
Juglans regia, presence of some genus of fungi; Aspergillus, Penicillium,Botryoderma,
Chaetomium, Phoma, Cladosporium and Pacilomyces in different intensities.
The two fungi Aspergillus and Penicillium appeared more dominants than others, therefore
they were chosen for the pathogenicity test. The results showed that the two species of fungi
preferred Juglans wood firstly were the size of infection was more than 10 times of any of the
other two woods. Eucalyptus showed similar response to that of Morus, but with Aspergillus
it was few bett
This study is conducted to identify the microbial content of some types of infant milk formula available in the local markets of the city of Baghdad and their conformity microbial limits sited by the Iraqi standard. Seventy samples were collected from trademarks of imported infant milk formula included of five samples of infant milk formula No (1) and five samples of follow-up formula No (2). These samples were collected randomly from shops in the local markets of Baghdad city on both sides of Karkh and Rusafa included the following kinds: Dialac 1, Dialac 2 ,Celia 1, Celia 2 ,Biomil 1, Biomil 2 , Nactalia 1, Nactalia 2, Novalac 1 , Novalac 2 , Similac 1 , imilac 2 , Guigos 1, Guigos 2. Some microbial tests were done which in
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Background: This research identified Streptococci spp. depending on culture, biochemistry, the VITEK technique, ability to produce biofilms, and antibiotic resistance. Aim: The goal of this study was to perform microbiological procedures to evaluate the qualitative qualities of mozzarella cheese against infective Streptococci using microbiological care. Methods: Sixty (60) mozzarella cheese samples were brought from diverse markets in Baghdad from October 2023 to December 2023 at the Zoonoses Research Unit and Veterinary Public Health Department, Veterinary Medicine College, University of Baghdad. Culture of samples on agar (MacConkey and blood) and aerobically incubated at 37°C for 48 hours. Gram staining purified colonies to
... Show MoreThis paper includes an experimental study of hydrogen mass flow rate and inlet hydrogen pressure effect on the fuel cell performance. Depending on the experimental results, a model of fuel cell based on artificial neural networks is proposed. A back propagation learning rule with the log-sigmoid activation function is adopted to construct neural networks model. Experimental data resulting from 36 fuel cell tests are used as a learning data. The hydrogen mass flow rate, applied load and inlet hydrogen pressure are inputs to fuel cell model, while the current and voltage are outputs. Proposed model could successfully predict the fuel cell performance in good agreement with actual data. This work is extended to developed fuel cell feedback
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