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.
Normally, bacteria exposed to antibiotics at sub minimal inhibitory concentrations (MIC) inside the host. Therefore, the current study aimed to comprehend the association among hemolysins, biofilm, as well as gentamicin resistance in local MRSA isolates. Around 35 Staphylococcus aureus locally isolated from different clinical specimens were employed in this study. Methicillin resistance was detected via cefoxitin disk diffusion and mecA amplification methods. MIC of gentamicin was estimated by broth microdilution method. Hemolysin genes involving hla, hlb, hld, and hlg were determined using multiplex polymerase chain reaction (PCR) technique. Microtiter plate method was employed for biofilm assessment in the presence and absence of gentamic
... Show MoreForeign direct investment (FDI) has been viewed as a power affecting economic growth (EG) directly and indirectly during the past few decades. This paper reviewed an amount of researches examining the relationships between FDI and EG, especially the effects of FDI on EG, from 1994 up to 2012. The results show that the main finding of the FDI-EG relation is significantly positive, but in some cases it is negative or even null. And within the relation, there exist several influencing factors such as the adequate levels of human capital, the well-developed financial markets, the complementarity between domestic and foreign investment and the open trade regimes, etc.
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The seasonal behavior of the light curve for selected star SS UMI and EXDRA during outburst cycle is studied. This behavior describes maximum temperature of outburst in dwarf nova. The raw data has been mathematically modeled by fitting Gaussian function based on the full width of the half maximum and the maximum value of the Gaussian. The results of this modeling describe the value of temperature of the dwarf novae star system leading to identify the type of elements that each dwarf nova consisted of.
The aim of this study is to uncover the views of female students of Shaqra University towards midterm tests. A survey of (42) items distributed in three fields. The study has been applied to a random sample of (421) students from two faculties. The results showed that all three fields have achieved an intermediate level. The results also indicated that there were no statistically significant differences in students’ views of midterm tests due to the specialization or academic level variables.
N, N′- bis[4-hydroxy phenyl] pyromillitdiimide [II] was prepared from the corresponding diamic acid , which was transfered to its new ester by the reaction with chloroethyl acetate [III ], [III] was used to prepare the novel hydrazide derivative [IV] , which was allowed to react with several aldehydes to yield the hydrazones [V – IX]. All the new compounds were synthesized , and characterized by their melting points .HNMR for some of them1FTIR,C,H,N analysis and ,
HTH Ali Tarik Abdulwahid , Ahmed Dheyaa Al-Obaidi , Mustafa Najah Al-Obaidi, eNeurologicalSci, 2023
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The effect of linear thermal stratification in stable stationary ambient fluid on free convective flow of a viscous incompressible fluid along a plane wall is numerically investigated in the present work. The governing equations of continuity, momentum and energy are solved numerically using finite difference method with Alternating Direct implicit Scheme. The velocity, temperature distributions
and the Nusselt number are discussed numerically for various values of physical parameters and presented through graphs. ANSYS program also used to solve the problem. The results show that the effect of stratification parameter is marginalized with the increase in Prandtl number, and the increase in Grashof number does not practically vary the
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Neoechinorhynchus iraqensis sp. n.(Acanthocephala: Neoechinorhynchidae) from the Freshwater Mullet, Liza abu.