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.
LK Abood, RA Ali, M Maliki, International Journal of Science and Research, 2015 - Cited by 2
Cancer remains a leading cause of mortality and morbidity worldwide. Advances in cancer therapies—including immunotherapies (e.g., checkpoint inhibitors, gene-targeted therapies), antibody-based cancer toxins, chemotherapy, radiotherapy, and surgery—have significantly improved survival rates 1,2. However, this progress has led to a surge in the prevalence of cardiovascular disease (CVD) among cancer survivors, now recognised as a leading cause of mortality in this population 3,4. These intersecting burdens highlight the growing need to prevent, detect, and manage cardiovascular complications in cancer care pathways and call for important initiatives in establishing cardio-oncology services globally.
The study included the collection of 75 bronchial wash samples from patients suspected to have lung cancer. These samples were subjected to a diagnostic cytological study to detect the dominant type of lung cancer. It was noticed that 33 patients proved to have a lung cancer out of 75 (44%) of these, 19 cases (57.6%)were diagnosed having Squamus cell carcinoma,7cases (21.21%) showed Adenocarcinoma ,6 cases (18.18%) were having small cell carcinoma while only one case (3.03%)was large cell carcinoma .Nearly 70% of cases were correlated with smokers .Bacteria were isolated from 53 patients in which 33 isolates were associated with the cancer cases while 20 of them from non infected patients. By using different morphological ,biochemical test
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S a mples of compact magnesia and alumina were evaporated
using CO2-laser .The
Processed powders were characterized by electron microscopy
and both scanning and transmission electron microscope. The results
indicated that the particle size for both powders have reduced largely
to 0.003 nm and 0.07 nm for MgO and Al2O3, with increasing in
shape sphericity.
Translation is both a social and cultural phenomenon, it can neither exist outside a social community and it is within society, nor it can be viewed as a medium of cross-cultural fertilization. This paper aims to investigate the difficulties that a translator may face when dealing with legal texts such as marriage and divorce contracts. These difficulties can be classified according to the present paper into syntactic, semantic, and cultural. The syntactic difficulties include word order, syntactic arrangement, unusual sentence structure, the use of model verbs in English, and difference in legal system. As to the semantic difficulties, they involve lack of established terminology, finding functional and lexical equivalence, word for word t
... Show MoreFever is a common illness in the pediatric age group ,the causes could be viral ,bacterial and fungal , this study was focused on bacterial pathogens as gram positive like Staphylococci, coagulase positive or negative ,Streptococci and gram negative like E-coli , Klebsialla ,Proteus, Pseudomonas, Burkhoderia , Acinetobacter and others like Pusturella ,E-alkalescendiaper, Haemophillus influenza and yeast like candida . Four thousand and seventy eight blood samples (4078)were collected in a period between January 2011 and the end of May 2012 at the child welfare hospital ,all the samples were cultured on suitable culture media and then biochemical tests were done using API-E 20 and sugar fermentation tests ,sensitivity
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Removal of solar brown and direct black dyes by coagulation with two aluminum based
coagulants was conducted. The main objective is to examine the efficiency of these
coagulants in the treatment of dye polluted water discharged from Al-Kadhymia Textile
Company (Baghdad-Iraq). The performance of these coagulants was investigated through
jar test by comparing dye percent removal at different wastewater pH, coagulant dose,
and initial dye concentration. Results show that alum works better than PAC under acidic
media (5-6) and PAC works better under basic media (7-8) in the removal of both solar
brown and direct black dyes. Higher doses of PAC were required to achieve the
maximum removal efficiency under optimum pH co
Background: Ambulatory peritoneal dialysis
introduced by Popvich et al (13) in 1978 , consists of a
four to five hours lavage of peritoneal cavity with 2000
ml of glucose solution .It remains a useful method for
treating patients with end stage renal failure till renal
transplantation becomes possible.
Objectives: The aim of the study is to evaluate the
value of cytological changes of mesothelial cells in
dialysate patients.
Methods: Within one year period, 32 cytological
peritoneal fluid samples were collected from patients
with end stage renal failure regardless of the underlying
causes, admitted to the dialyzing unit in Kadimya
Teaching Hospital. Smears were prepared and fixed in
95 % ethyl al
In this work, an experimental analysis is made to predict the thermal performance of the natural-convection phenomenon from a heated vertical externally finned-tube to surrounding air through an open-ended enclosure. Two different configurations of longitudinal rectangular fin namely, continuous and interrupted are utilized with constant thickness, different numbers, and different heights are extended radially on the outer surface of a heated tube. The tube is heated electrically from inner surface with five varied power input magnitudes. The effect of fins configuration, fins number, fins height, and heat flux of the inner tube surface on the thermal performance of natural c
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Phenomena of an abnormal genitalia was among some specimens of Cicindela aulica Dej.
Collected from Iraq. The fore tarsi of male were asymmetrical having its basal three segments
dilated and clothed beneath with fine bristles as in normal male. While those of the right leg
were found simple as in normal females. Dissection of the genialia of these specimens
showed that they were of two types of both male and female structures.