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.
Background: Toxoplasmosis is a very common infection caused by the obligate intracellular protozoan parasite. This parasite is called Toxoplasma gondii widely distributed around the world . Toxoplasma gondii can be vertically transmitted to the fetus during pregnancy and may cause wide range of clinical manifestations in the offspring.
Objective: To determine seroprevalence Immunoglobulin G (IgG) and Immunoglobulin M (IgM ) to toxoplasma gondii among pregnant women and to identify the risk factors.
Type of the study: A cross-sectional study.
Methods: A total of 110 blood samples of pregnant women were collected from
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In the present work is the deposition of copper oxide using the pulsed laser deposition technique using Reactive Pulsed Laser as a Deposition technique (RPLD), 1.064μm, 7 nsec Q-switch Nd-YAG laser with 400 mJ/cm2 laser energy’s has been used to ablated high purity cupper target and deposited on the porous silicon substrates recorded and study the effect of rapid thermal annealing on the structural characteristics, morphological, electrical characteristics and properties of the solar cell. Results of AFM likelihood of improved absorption, thereby reducing the reflection compared with crystalline silicon surface. The results showed the characteristics of the solar cell and a clear improvement in the efficiency of the solar cell in the
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Objective(s): To measure serum C-reactive protein (CRP) titer as a predictive diagnosis of acute hepatitis C virus (HCV)
infection.
Methodology: Two hundred and ten patients with acute HCV infection and 234 apparently healthy individuals as
control group were enrolled in this study in Baghdad medical city (Teaching Laboratories). The patents include
74(35.2%) females and 136 (64.8%) males with mean age (27±16.5) years. The control group includes 114 (48.7%)
females and 120 (51.3%) males with mean age (26±5.8) years. Blood samples were collected from out patients from
Alfadul in Baghdad city. Sera were separated and stored at 20 0
C. The diagnosis of acute HCV infection was based on
detection of HC Ag and anti- H
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The design of this paper is to find the possible correlation of Epstein Barr virus infection ina group of Iraqi women with cervical carcinoma though detection of Latent Membrane Protein 1 (LMP1) in these cervical tissues. Paraffinized blocks of two groups were included. The first sample of 30 cervical carcinomatous tissues and 15 biopsies from an apparently normal cervical tissues. All the samples were sectioned on a positive charged slides with 4 mm – thickness then submitted for immunohistochemical (IHC) staining to detect viral LMP1 expression. Sixty three percentage (19 out of 30) of the studies group showed positive overexpression as shown in with a significant association of the expression with cervical cancer with a significant ass
... Show MoreBackground: Whey protein is the green-yellow colored, liquid portion of the milk, and it is also called the cheese serum, it is obtained after the separation of curd, during the coagulation of the milk. It contains a considerable amount of α-helix pattern with an evenly distributed hydrophobic and hydrophilic as well as basic and acidic amino acids along with their polypeptide chain. The major whey protein constituents include β-lactoglobulin (β-LG),α-lactalbumin (α-LA), immunoglobulins (IG), bovine serum albumin (BSA), bovine lactoperoxidase (LP), bovine lactoferrin (BLF) and minor amounts of a glycol macro peptide (GMP). Osseointegration can be defined as a process that is immune driven which leads to the formatio
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