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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Low-pressure capacitively coupled RF discharge Ar plasma has been studied using Langmuir probe. The electron temperature, electron density and Debay length were calculated under different pressures and electrode gap. In this work the RF Langmuir probe is designed using 4MHz filter as compensation circuit and I-V probe characteristic have been investigated. The pressure varied from 0.07 mbar to 0.1 mbar while electrode gap varied from 2-5 cm. The plasma was generated using power supply at 4MHz frequency with power 300 W. The flowmeter is used to control Argon gas flow in the range of 600 standard cubic centimeters per minute (sccm). The electron temperature drops slowly with pressure and it's gradually decreased when expanding the electro
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Water samples were collected from output of water for Al-Wahda plant where located in al-karrada area in Baghdad city to study water contamination with bacteria, fungi and Algae. The study lasted one year started on August, 2016 to July,2017.Results were acquired according to two tests performed, the first is biological test included total coliform,E.coli, pseudomonas aeruginosa, total fungi, Diatom and non Diatom Algae and the second is physiochemical test included temperature, turbidity and residual chlorine. The results of bacteria were within the permitted specification in the Iraqi standards no. 14/2270 for the year 2015 except August was exceeded the permitted standard for total coliform, it was 1.1< cell/100 ml.Total Fungi, Dia
... Show MoreThis study was aimed to study the effect of adding transglutaminase (TGase) on the mechanical and reservation properties of the edible films manufactured from soybean meal protein isolate (SPI) and whey protein isolate(WPI). The results showed an improvement in the properties with increase in the WPI ratios. Thickness of the SPI films amounted 0.097 mm decreased to 0.096 mm for the WPI: SPI films at a ratio of 2:1, when TGase was added decreased to 0.075 mm. While the tensile strength increased from 7.64 MPa for SPI films to eight MPa for the WPI: SPI films at a ratio of 2:1, when TGase was added increased to 11.04 MPa. Also, the elongation of the WPI: SPI films at a ratio of 2:1 presence of the TGase decreased to 40.6% compared wit
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In this research, the size strain plot method was used to estimate the particle size and lattice strain of CaTiO3 nanoparticles. The SSP method was developed to calculate new variables, namely stress, and strain energy, and the results were crystallite size (44.7181794 nm) lattice strain (0.001211), This method has been modified to calculate new variables such as stress and its value (184.3046308X10-3Mpa) and strain energy and its value (1.115833287X10-6 KJm-3).
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Proteus mirabilis ? -lactamase of local isolates number 4TF represent karkh side and 20TF represent rusafa side of Baghdad were extracted and purified 23.17, 25.23 fold with yield of 36.66 %, 37.5% and specific activity 11.8, 12.6 of unit/ mg protein by DEAE –cellulose and Sepharose 4B (respectively ).Molecular weight of both enzyme was about 35500 Dalton determined by gel filtration. The study indicated that the isoelectric point of purified ? -lactamase that extracted from isolate number 4TF and 20TF was 5.4.
This study was aimed to determine the mutations and single nucleotide polymorphisms (SNPs) in exon 3 and 7 of estrogen receptor beta (ESR2) gene in women with breast cancer from Iraq. Different samples (blood, fresh tissue with blood from same patient, and formalin fixed paraffin embedded, FFPE) were collected from women with breast cancer. Molecular analysis exon 3 and 7 in ESR2 has been studied by using PCR. It was found exon 3 and 7 in ESR2 were revealed as a single band with size 151 and 157 bp, respectively. There was no SNP in exon 3 has been identified. While three novel polymorphisms (ACT, AGG and GCA) were detected in exon 7, the type of those polymorphisms deletion for ACT and AGG while substitution polymorphism for GCA. From this
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The main idea of this paper is to define other types of a fuzzy local function and study the advantages and differences between them in addition to discussing some definitions of finding new fuzzy topologies. Also in this research, a new type of fuzzy closure has been defined, where the relation between the new type and different types of fuzzy local function has been studied
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In this research, the influence of the fermentation treatments and baking in Iraqi’s flour type (Ibaa 99, Al-rashed, Tamus, Abu-grabe) and Turkish flour type (Muaamel) on phytic acid was investigated. In whole wheat flour, the phytic acid was (1500, 1290, 1450, 1230, 1440 ( mg/ 100 g flour respectively, and the inorganic phosphorous was (29.18, 25.15, 23.89, 20.85, 22.83) mg/100 g flour respectively. The dough prepared from flour with a higher phytic acid content also contained higher amount of phytic acid. During fermentation, degradation of phytic acid occurred. The cumulative loss of phytic acid after fermentation in all type of dough was ~ 23, 22, 34, 26 and 27% respectively،While increased of inorganic phosphorous occurred. The c
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