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.
Abstract:
This investigation was carried out to study the nutritional adequacy for
infant milk formula, which imported by Iraqi Ministry of Trade, and are
available in local markets .Most of these formulas contained nearly the same
composition of nutrients which are ,Matines ,Sunny Boy , Salsabeel AL- Badie
,Moroug, ,Charton ,Materna Lery Celia ,Lacstar Lailac,Nactalia. yet they are
unbalanced for providing the daily nutritional requirements for infants whom
depend on bottle feeding for six times daily in their first six month of age. As
there were an increase in daily intake for protein content and most vitamins
that included D, E, C, B1, B2, Niacin, B6, B12, and Biotin as well as most
minerals namely Calci
The study aimed to estimate the content of lead and determine the quality of the internal coating of metal cans through electrical conductivity as well as to determine the accuracy of the information card for some types of canned food that available in local markets. The information card test showed that all of these samples contained the name of the food, trade mark, country origin, weight, and components, as was indicated by the company producing in all of them except for the C12 sample which was otherwise, and the batch number was mentioned in all samples except for the C3 and C17 which was not clear and not mentioned in the C21, and the validity period was observed (produce and fini
... Show MoreOne of these plants utilized in traditional medicine is Lactuca seriolla Linn., which belongs to the Asteraceae family. It goes by a variety of common names in the world, including prickly lettuce, wild lettuce, jagged lettuce, and Kahu and khas. The work aimed to isolate and characterize some bioactive constituent(s) from the aerial part of Lactuca serriola utilizing Combiflash NEXTGEN and high-performance liquid chromatography (HPLC). Lactuca serriola (aerial part) was extracted with 80% ethanol, then fractionated with hexane. Then 250 mg of hexane extract was mixed with 4 g of silica gel and loaded in cartilage, then bounded to the gold column of combi flash using a solvent system comprised of ethyl acetat
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Aluminum doped zinc selenide ZnSe/n-Si thin films of (250∓20 nm) thickness with (0.01, 0.02 and 0.03), are depositing on the two type of substrate (glass and n-Si) to manufacture (ZnSe/n-Si) solar cell through using thermal vacuum evaporation procedure. physical and optoelectronic properties were examined for the samples. X-Ray and AFM techniques are using to study the structure properties. The energy band gap of as-deposited ZnSe thin films for changed dopant ratio were ranging from (2.6-2.68 eV). The results of Hall effect show that pure and doping films were (p-type), and the concentration carriers and the carriers mobility increases with increase Al-dopant ratio. The (C-V) have shown that the heterojunction were of abrupt type. In add
... Show MoreThis study was conducted to investigate the effect of feeding diets containing different levels of parsley on the hematological traits of local Iraqi geese. A total of twenty-four local geese, one year old, were used in this experiment. The birds were allocated into four treatment groups, consisting of six geese each. Treatment groups were: control diet (C) (free from parsley); T1: control diet + 80 g/d parsley; T2: control diet + 160 g/d parsley; and T3: control diet + 240 g/d parsley. At the end of the experiment, blood samples were obtained from all geese from the brachial vein by venipuncture. Hematological traits included in this study were red blood cells count (RBC), hemoglobin concentration (Hb), packed cell volume (PCV), mean cell
... Show MoreIn this paper, we establish the conditions of the occurrence of the local bifurcations, such as saddle node, transcritical and pitchfork, of all equilibrium points of an eco-epidemiological model consisting of a prey-predator model with SI (susceptible-infected) epidemic diseases in prey population only and a refuge-stage structure in the predators. It is observed that there is a transcritical bifurcation near the axial and free predator equilibrium points, near disease-free equilibrium point is a saddle-node bifurcation and near positive (coexistence) equilibrium point is a saddle-node bifurcation, a transcritical bifurcation and a pitchfork bifurcation. Further investigations for Hopf bifurcation near coexistence equilibrium point
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In current study a computation fluid dynamic (CFD) technique was used to investigate the effect of groynes shape and spacing on the scour pattern and the maximum scour depth in open channel flow. CFD model have been validated throughout comparing the numerical results with three previous experimental studies for a single groyne located in open channel with three different shapes (L, quadrant, and parabola shapes). The comparison revealed very good agreement between numerical results of the maximum scour depth with the results of all experimental models. Moreover, investigations of the effect of multi-groynes (three groynes and four groynes) arranged in parallel with constant spacing and also with variable spacing have been done, the
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Objective: Many studies focused on clinical cases such as ovariohysterectomy of bitches and scarcely mentioned the histological features. The present study describes the cytoarchitecture characteristics of a local dog’s mature adult reproductive tract. Materials and Methods: Sixteen samples of uterus and cervix were obtained from local breed bitches to conduct this study. The organs were processed according to routine histopathological protocol and stained with hematoxylin and eosin, Masson’s trichrome, and combined Alcian blue (2.5 pH) and PAS (AB-PAS) stains. Results: The mature endometrium formed numerous short epithelial folds and epithelial crypts composed of mucous cells and cuboidal cells. The core of the endometrium is c
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This study was conducted at the poultry farm of the Department of Animal Production/College of Agriculture/University of Baghdad/Abu Ghraib, on 252 birds (180 females and 72 males). This study aims to observe the effect of melatonin implantation and exposure to different light colors and their interaction on characteristics of fertility and hatching of local Iraqi chickens. The birds were divided into three sections (white, red and green) each section contains two lines, one of which has been planted melatonin under the skin of the neck of birds and the other has not been planted hormones. The results showed that melatonin implantation and exposure to different light colors did not significantly affect the hatching rate of fertilized eggs a
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