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 MoreThe current study included measuring the percent of protein in the extract of nematode Ascaridia galli that infect chickens, it was 1.157% and equivalent to 11.570 mg /L., as well as the amino acid analysis in the nematode A.galli by using a high-performance liquid chromatography technique (HPLC), as was detect five types for amino acids in this extract Leucine, Threonin, Serine, Methionine and Valine as the amount of these amino acids in the extract was as follows 132.973, 26.994, 10.453, 2.243 and 1.888 mg /L., respectively, and other amino acids which Glutamic, Histidine and Tyrosin did not exist in the nematode A.galli.
due to the presence of chemoresistance and the risk of tumor recurrence and metastasis. There is a pressing necessity to develop efficient treatments to improve response for treatment and increase prolong survival of breast cancer patients. Photodynamic therapy (PDT) has attracted interest for its features as a noninvasive and relatively selective cancer treatment. This method relies on light-activated photosensitizers that, upon absorbing light, generate reactive oxygen species (ROS) with powerful cell-killing outcomes. Nuclear factor kappa B (NF-κB), a transcription factor, plays a key role in cancer development by regulating cell proliferation, differentiation, and survival. Inhibiting NF-κB can sensitize tumor cells to chemotherapeuti
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Background: The diagnosis of prostatic pathology may be of challenging , as some difficult and suspected, atypical cases may lack basal cell layer by routine H&E sections . Antibodies against 34BE12(HMW-CK) and p63 aid the diagnosis of such cases , to distinguish benign from malignant prostatic lesions.
Objective: to identify basal cells in atypical prostatic lesions ,and distinguish benign from malignant prostatic lesions.
Type of the study: A retro-spective study.
Methods: 115cases of paraffin embedded prostatic tissue blocks ,diagnosed as : 76 cases were benign prostatic hy
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In this research, we studied the effect of concentration carriers on the efficiency of the N749-TiO2 heterogeneous solar cell based on quantum electron transfer theory using a donor-acceptor scenario. The photoelectric properties of the N749-TiO2 interfaces in dye sensitized solar cells DSSCs are calculated using the J-V curves. For the 〖(CH_3)〗_3 COH solvent, the N749-TiO2 heterogeneous solar cell shows that the concentration carrier together with the strength coupling are the main factors affecting the current density, fill factor and efficiency. The current density and current increase as the concentration increases and the strength coupling increases as the N749-TiO2 heterogeneous in solar cell. However, the efficiency is more sens
... Show MoreThis experiment was conducted to study the effect of feeding diets containing different levels of parsley on the blood biochemical characteristics of local Iraqi geese. A total of twenty-four local geese, one year old, were used in this experiment during the period from the beginning of October to the end of December. The birds were allocated for Four treatment groups consisted of six geese each. Treatment groups were as follows: Control diet (T1) (free from parsley), T2: Control diet + 80 g / d parsley, T3: Control diet + 160 g / d parsley; T4: Control diet + 240 g / d parsley. At the end of experiment, blood samples were obtained from all geese by venipuncture from brachial vein and blood plasma samples were prepared. Blood biochemical tr
... 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 MoreObjective: 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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In 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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