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.
يعد هذا النص أحد النصوص المسمارية المصادرة التي بحوزة المتحف العراقي، ويحمل الرقم المتحفي (235869)، قياساته )12،7x 6x 2،5سم). يتضمن مدخولات كميات من الشعير،أرخ النص الى عصر أور الثالثة (2012-2004 ق.م) و يعود الى السنة الثالثة من حكم الملك أبي-سين (2028-2004 ق.م)،أن الشخصية الرئيسة في هذا النص هو)با-اَ-كا مسمن الماشية( من مدينة أري-ساكرك، ومقارنته مع النصوص المسمارية المنشورة التي تعود الى أرشيفه يبلغ عددها (196) نصاً تضمنت نشاطاته م
... Show MoreS 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.
This study aimed to assess the efficiency of Nerium oleander in removing three different metals (Cd, Cu, and Ni) from simulated wastewater using horizontal subsurface flow constructed wetland (HSSF-CW) system. The HSSF-CW pilot scale was operated at two hydraulic retention times (HRTs) of 4 and 7 days, filled with a substrate layer of sand and gravel. The results indicated that the HSSF-CW had high removal efficiency of Cd and Cu. A higher HRT (7 days) resulted in greater removal efficiency reaching up to (99.3% Cd, 99.5% Cu, 86.3% Ni) compared to 4 days. The substrate played a significant role in removal of metals due to adsorption and precipitation. The N. oleander plant also showed a good tolerance to the uptake of Cd, Cu, and Ni ions fr
... Show MoreThis study assesses the short-term and long-term interactions between firm performance, financial education and political instability in the case of Malaysia Small to Medium Enterprises (SMEs). The simultaneous insertion of financial education and political instability within the study is done intentionally to inspect the effect of these two elements in one equation for the Malaysian economy. Using the bound testing methodology for cointegration and error correction models, advanced within an autoregressive distributed lag (ARDL) framework, we examine whether a long-run equilibrium connection survives between firm performance and the above mentioned independent variables. Using this method, we uncover evidence of a positive long-term link b
... Show MoreMelanoidins can be diagnosed using the Fourier transform infrared (FTIR) technique. UV/Vis is an effective tool for both qualitative and quantitative analysis of chemical components in melanoidin polymers. The structural and vibrational features of melanoidin synthesized from D-glucose and D-fructose are identical, according to FTIR spectra, with the only difference being the intensity of bands. Using FTIR spectra, the skeleton of melanoidin is divided into seven major regions. The existence of the C=C, C=N, and C=O groups in all melanoidins formed from fructose and glucose with ammonia is confirmed by the areas ranging from 1600 to 1690 cm-1, and the band is largely evident as a broad shoulder. Both melan
... Show MoreCompassion competence is a nurse's ability to provide patient-centered care and communicate with patients in a sensitive and insightful manner. This descriptive cross-sectional survey study aimed to assess the compassion competence of a multinational group of nursing students.
A total of 1,158 undergraduate nursing students participated in this study, and stratified random sampling method was used to select participants from the 100 study level to the 400 or 500 level. Data were collected using the Compassion Competence Scale.
<Using remote sensing technology and modeling methodologies to monitor changes in land surface temperature (LST) and urban heat islands (UHI) has become an essential reference for making decisions on sustainable land use. This study estimates LST and UHI in Salah al-din Province to contribute to land management, Urban planning, or climate resilience in the region; as a result of environmental changes in recent years, LANDSAT Satellite Imagery from 2014- 2024 was implemented to estimate the LST and UHI indexes in Salah al-din Province, ArcGIS 10.7 was use to calculate the indices, and The normalized mean vegetation index (NDVI) was calculated as it is closely related to extracting (LST