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.
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ABSTRACT
The study was conducted at the ruminant research station of the general commission for agricultural research/Ministry of Agriculture, as well as the laboratory of genetic resources of the department of livestock/Ministry of Agriculture and the laboratory of the college of agriculture engineering science, with the aim of determine the genotypic of the expression region (intron 2 and part of exon 3) of the LHX3 gene And its relationship to the fertility rate in local and Shami goats. For this purpose, the RFLP technique was used, and the percentages of genotypes for the LHX3 gene in the local goat sample were 29.17, 50.00, 20.83 for the TT, AT, and AA genotypes, respectively, while in the Shami goa
... Show MoreThe aim of this study to investigate the microbial load and type of preservative for the types of yogurt available in the Iraqi market to ensure the safety of food provided to the consumer and protect through examining the types of yogurt from harmful bacteria as well as to contain ratios acceptable to yeasts and molds is to find out by comparing models curd careless Iraqi standard quality(ISQ) and see how they conform to these specifications have been collecting 12 brands of yoghurt types it was been (Kala, Activia 1, Activia 2, Mazia, Shelan, Aib, Mersin, Morsi, Al-Safi, Zabady, Zakho, Arbil). Bacteriological tests were conducted on samples of yogurt (total bacterial count, coliform count, counting yeasts and molds). The results showed
... Show MoreAs part of our research on efficiency improvement of PERC (Passivated Emitter Rear Solar Cell), achieving very low reflectivity values of solar cell surface is a must. One of the most advance technologies to do so is the use of advanced texturing for the front surface of the cells. This texture, also known as Black Silicon, consists of peaks and valleys of nano metric dimensions and capable of dramatically reducing the reflectance of the front surface. A reflectance around 5% was reached ,using simulation, when using a Black-Silicon texturing with height of 50nm with peak rounding of 5nm. Even though this texturing may affect other parameters such as series resistance or surface recombination, as a starting point
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To limit or reduce common microbial contamination occurrence in dairy products in general and in soft cheese in particular, produced in locally plants, this study was performed to demonstrate the possibility of implementing HACCP in one of dairy plants in Baghdad city
HACCP plan was proposed in soft cheese production line. A pre-evaluation was performed in soft cheese line production, HACCP Pre-requisites programs was evaluated from its presence and effectiveness. The evaluation was demonstrated risk in each of: Good Manufacturing Practice (GMP) program, evaluated as microbial and physical risk and considered as critical r
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The study aimed to evaluate the information label of some local pickle products and estimate sodium benzoate therein. 85 samples of locally made pickles were collected from Baghdad city markets and randomly from five different areas in Baghdad it included (Al-Shula, Al-Bayaa, Al-Nahrawan, Al-Taji, and Abu Ghraib), which were divided into groups P1, P2, P3, P4 and P5, respectively, according to those areas, samples information label was scanned and compared with the Iraqi standard specification for the information card of packaged and canned food IQS 230, the results showed that 25.9% of the samples were devoid of the indication card informa
... Show Morehe study aimed to purify of Leucine aminopeptidase (LAP) from sera of hyperthyroidism patients and its relation to some thyroid hormones (TSH, T 3 , T 4 ) of subclinical hyperthyroidism and hyperthyroidism patients with lipid peroxidation levels that may be play a role in this diseases. Specimens were collected during the time from Nov 2017 until Jan 2018 from Endocrine and Diabetes Center, blood samples were collected from fifty healthy control and one hundred patients, patients were divided into two groups consisted of (50) with hyperthyroidism and (50) with subclinical hyperthyroidism. The aged for all subjects ranged (15-60) years with body mass ranged ((25- 29) kg/m 2 . The purification is done by addition of ammonium sulfate, dial
... Show MoreThis paper deals with the blow-up properties of positive solutions to a parabolic system of two heat equations, defined on a ball in associated with coupled Neumann boundary conditions of exponential type. The upper bounds of blow-up rate estimates are derived. Moreover, it is proved that the blow-up in this problem can only occur on the boundary.
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