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.
The present study examines the extraction of lead (Pb), cadmium (Cd) and nickel (Ni) from a contaminated soil by washing process. Ethylenediaminetetraacetic acid disodium salt (Na2EDTA) and hydrochloric acid (HCl) solution were used as extractants. Soil washing is one of the most suitable in-situ/ ex-situ remediation method in removing heavy metals. Soil was artificially contaminated with 500 mg/kg (Pb , Cd and Ni ). A set of batch experiments were carried out at different conditions of extractant concentration , contact time, pH and agitation speed. The results showed that the maximum removal efficiencies of (Cd, Pb and Ni ) were (97, 88 and 24 )&nbs
... Show MoreThe adsorption of Congo red (CR) dye on modified synthetic zeolite 5A , the general name of which is Linde Type A (LTA)which is modified by amino mercepto thiadiazole (AMT) and have been characterized by using fourier transform infrared (FT-IR) , x-ray diffraction (XRD) spectroscopies, atomic force microscopy (AFM) and scanning electron microscope (SEM) analysis.In this work Modified zeolite was utilized as adsorbent to remove (CR) dye from aqueous solution by adsorption. Batch experiments were conducted to study the effects contact time , initial concentration of adsorbate and temperature on dye adsorption. The equilibrium adsorption data were analyzed by using several isotherm models ( Freu
... Show MoreIn this study miconazole nitrate was formulated as topically applied emulgel; different formulas were prepared using sodium carboxymethylcellulose (SCMC) and carboxypolymethylene (carbomer 941) as gelling agents. The influence of type of gelling agent and concentration of both oil phase and emulsifying agent on drug release was studied and compared with commercially available miconazole nitrate cream (Mecozalen®). The results of in vitro release showed that SCMC emulgel bases gave better release than carbomer 941 bases and the release of drug increase from both bases as a function of increasing the concentration of emulisifying agent. The oil phase had retardation effect when
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There are numerous ways to define the traits of influential professors, including behavior (such as warmth, civility, and clarity), knowledge (of subject matter and students), and beliefs, among many others. According to research; influential teaching can also defined as the capacity to improve student achievement. This; is only one way to express effectiveness, as stated. However; the preferred definition of high-quality teaching in the United States and many other countries is teacher effects on student achievement. Therefore; this study aimed to determine the characteristics of influential professors in Online Teaching at the English department. This study; believed to be significant for college professors who will show them how
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The presence of hydrocarbons in the soil is considered one of the main problems of pollution. In our current study, eight samples isolated from soil saturated with hydrocarbons were taken from different areas of Baghdad, Iraq. In this study, 5 isolates belonging to Pseudomonas aeruginosa by 99%, 4 isolates to Klebsiella pneumoniae by 98%, and 3 isolates to Enterobacter hormaechei by 97% were diagnosed in different ways. A molecular examination was also conducted by 16sRNA. We recorded P. aeruginosa, K. Pneumoniae and E. hormaechei as new local isolates in NCBI. In addition, a comparison was made between our isolates and the global isolates to determine the degree of convergence in the evolutionary line. The genes alkB and nahAc7 were diagno
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The presence of residual antibiotics in water results in the development of antibiotics resistant genes. The available wastewater treatment systems are not capable of removing such antibiotics from sewage. Thus, antibiotics need to be removed before the discharge of wastewater. Adsorption is among the promising techniques for the wastewater treatment to aid the removal of a wide range of organic and inorganic pollutants. The present work is a contribution to the search for an economical method for the removal of low concentrations of amoxicillin (AMX) from water by adsorption on water treatment residue, WTR, taken from a local drinking water facility. The chemical composition and the adsorptive characteristics of the material were first
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The study aimed to determine the impact of energy for the north and south magnetic poles on the the growth of bacteria isolated from cases of tooth decay, 68 swabs were collected from surfaces of faulty tooth, the detected of Staphylococcus aureus
... Show MoreIn this study, from a total of 856 mastitis cases in lactating ewes, only 34 Streptococcus agalactiae isolates showed various types of resistance to three types of antibiotics (Penicillin, Erythromycin and Tetracycline). St. agalactiae isolates were identified according to the standard methods, including a new suggested technique called specific Chromogenic agar. It was found that antibiotic bacterial resistance was clearly identified by using MIC-microplate assay (dilution method). Also, by real-time PCR technique, it was determined that there were three antibiotics genes resistance ( pbp2b, tetO and mefA ). The high percentage of isolate carried of a single gene which was the Tetracycline (20.59%) followed by percentage Penicillin was
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The present study provides a new insight into valuable information on the diverse structure of the Anisakid population and discusses the limited species richness in the Nemipterus japonicus (Bloch,1791) (Perciformes, Nemiperidae). The fishing area consists of various locations in the Arabian Gulf (29°58 0 33 00 N48°28 0 20 E). A total of 315 marine fish were examined, (n=287) were infected. Larval stages (n= 763) encysted within the mesenteries peritoneum and viscera of fish organs were isolated, with a prevalence of 91.11% of infection and, the intensity was 2.65. Molecular analysis was carried out on thirty individuals who have examined the morphology and showed some appearance differences, by amplifying internal transcribed spacers
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