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.
In this work, lanthanium (III) complexes were synthesized using by Schiff base ligand (L) derived from benzaldehyde and o-aminoaniline with five amino acids (AA) from glycine (Gly), L-alanine (Ala), L-valine (Val), L-asparagine (Asp) and DL- phenylalanine (Phe). The Schiff base ligand has been characterized by elemental analysis, (MASS, FTIR, 1HNMR, 13CNMR, UV-VIS) electronic spectra. The structures of the new complexes have been described of analysis of elements, molar conductivity, (UV-Vis electronic, FTIR, mass) spectra also magnetic moment. The molar conductivity values of the complexes indicat this every of complexes are electrolytes and other analytical studies reveal octahedral geometry for La (III) ion. The Schiff base ligand, five
... Show MoreEscherichia coli has been recognized worldwide as the most common causative agent for severe infections of the urinary tract. Colibactin is a genotoxin produced through a gene cluster called polyketide synthase (pks) island by members of Enterobacteriaceae. Limited information is available about the frequency of colibactin in E. coli isolates in Iraq. Hence, this study aimed to examine the frequency of some colibactin genes (CIbA and CIbQ) in clinical isolates of E. coli obtained from urinary tract infections (UTIs) in Iraq. Between October 2023 and January 2024, 120 urine samples were collected from females diagnosed with UTIs in Iraqi hospitals. 70 E. coli isolates were isolated after identification by biochemical methods and confirmed by
... Show MoreEuphemisms are advantageous in people’s social life by turning sensitive into a more acceptable ones so that resentful feelings and embarrassment can be avoided. This study investigates the ability of Iraqi English learners in using euphemistic expressions, meanwhile, raising their awareness and the faculty members in English teaching faculties regarding the relevance of discussing the topics that demand euphemisation. This study comprised three stages: initial test, explicit instruction with activities, and a final test for the students’ development in this domain. A test has been distributed among 50 respondents, who are at the fourth year of their undergraduate study at the University of Babylon/ College of Basic Education. The lo
... Show More
(64)
(65)
(4)
(4)
Background: Insufficient sleep due to excessive media use is linked to decrease physical activity, poor nutrition, obesity, and decreased overall health-related quality of life.
Objectives: To assess the effect of using the internet and social media on the sleep of 4th-stage secondary school students.
Subjects and Methods: Cross-sectional study with the analytic element; for 500 secondary school students, obtained by choosing two schools randomly from each of the six educational directorates, by using a structured questionnaire.
Result: Secondary scho
... Show More
(4)
(2)
The ZnO nanoparticles were synthesized at various precursor concentrations i.e. 0.05, 0.1, and 0.5 M by biosynthesis method based on Pometia pinnata Leaf Extracts. Initial nanoparticle concentration influenced the optical bandgap, shape, and structure of nanoparticles. The photodegradation process was carried out under UV illumination. The efficiency of MB degradation was determined by measuring the decrease in MB concentration and by analyzing the optical absorption at 663 nm recorded by UV-Vis spectroscopy. Results showed that the biosynthesized ZnO nanoparticles exhibited efficient photodegradation of MB, with a maximum degradation rate of 80% after 90 minutes of exposure to UV-C light. The study highlights the potential of Pometia pi
... Show More
(7)
(2)
Three types of extracts ( aquatic, alcoholic, and oily ) were prepared from the fruits of coconuts, and a series of chemical tests were conducted in addition to the use of the FTIR equipment to determine the active locations in the prepared extracts. The results indicated the presence of active compounds (tannins, saponins, flavonoids, turbines and steroids) in the extracts prepared from the fruits of coconuts, also the antimicrobial capability of these extracts were tested on pathogenic bacteria isolated from wounds and burns infections cases. The results proved that the concentration 80 mg/ml of the aquatic extract is the minimum inhibitory concentration for the microbes: Proteus vulgaris and Pseudomonas fluorescence, while the
... Show More
This research examines how Al-Sabah newspaper addressed violations of women’s rights in Iraq. Using content analysis, a sample of 157 articles published from July 1, 2021, to December 31, 2021, was analyzed. The study reveals diverse journalistic treatments, with interpretive approaches employed in addressing violations of Iraqi women’s rights. Social violations of women’s rights were the primary focus, and various forms of newspaper articles, including opinion pieces, were commonly used.
(1)