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 appliance of milligauss meter was designed by Qusay Ismail to measure the induce of electromagnetic field for home appliance which are put at a distance from milligauss meter (15-30-60)cm .The results showed some appliance has recorded higher than normal acceptable level of electromagnetic radiation emissions and produced radiation of (350650)milligauss as for the rest of appliances has recorded values which are ranged between (1200)milligauss ,laptop was recorde radiation generally lower than from desktop and computer moniter (CRT).The radiation ,intensity decrease with increasing distance.
Human urinary Adenosine-3',5'-cyclic monophosphate (cAMP) was studied in 90 normal healthy volunteers (49 males and 41 females) aged between (11 months -55 years), and 86 leukemia patients (48 males and 38 females) of four types (25 ALL, 28 AML, 14 CLL, 19 CML) aged between (11 months - 65 years). The study includes the following:- Extraction and purification of urinary cAMP from the interfering nucleotides, proteins, phosphates and pyrophosphates, by using Zinc sulphate –Barium hydroxide precipitation. The extracted cAMP was purified by using Dowax 50W-H+ hydrogen form column chromatography (1x5 cm). Identification of the purified cAMP, this was achieved by applying the following techniques: a- U.V analysis: -
... Show MoreThis work was conducted to study the extraction of pelletierine sulphate from Punica granatum L. roots by liquid membrane techniques. Pelletierine sulphate is used widely in medicine. The general behavior of extraction process indicates that pelletierine conversion increased with increasing the number of stages and the discs rotation speed but high rotation speed was not favored because of the increased risk of droplet formation during the operation. The pH of feed and acceptor solution was also important. The results exhibit that the highest pelletierine conversion was obtained when using two stages, (10 rpm) discs speed of stainless steel discs, (pH=9.5) of feed solution and (pH=2) of acceptor solution in n-decane. Assuming the existen
... Show MoreResearchers are interested in the issue of children abuse and they look for its cause in the past and present. Their interest is limited to identifying penal liability which is caused by children abuse away from focusing on civil liability. So, the study is going to clarify the parents' responsibility for children abuse rather than civil liability of the medic in case he wouldn’t notify the authorities about the case according to the American law rules and the attitude of Iraqi law rather than some judicial application of civil cases that were exposed to American judiciary concerning children abuse.
The mucilage from the seeds of Lallemantia royleana family Labiatae was extracted and subjected to preformulation study for evaluation of its suitability for use as suspending agent. Furosemide suspensions were prepared using (1.5% w/v) of the extracted Lallemantia royleana mucilage, (1.5% w/v) chitosan and (0.35% w/v) xanthan gum. The mucilage was white in color and the average yield of dried mucilage obtained from L.royleana nutlets was 14 % w/w of the seeds used. It is sparingly soluble in water but swells in contact with it, giving a highly viscous solution. It is slightly acidic to neutral. It was found that the extracted natural mucilage of Lallemantia royleana exhibited a higher viscosity profil
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In this work, the nano particles of Na-A zeolite were synthesized by sol –gel method. The samples were characterized by X-ray diffraction (XRD), X-ray luorescence (XRF), Surface area and pore volume, Atomic Force Microscope (AFM) and Fourier Transform Infrared Spectroscopy (FTIR). Results show that the nano A zeolite is with average crystal size is 74.77 nm., Si/Al ratio 1.03, BET surface area was 581.211m2/g and the pore volume for NaA was found equal to 0.355cm3/g.
A study carried out on ceramic material made at (a-Al2O3) doped with MgO (0.5 , 0.3 , 0.2,0.1)%,with particle size at 63mm.
A Hydraulic press of 5kn at diameter of 2cm.A nnalelling at 1500Co and 6 hrs still to see the effect on the changes of the dielectric material. With frequency range at (1K – 1M) Hz. And the result show that at percentage of 0.5% of MgO, the real dielectric material decreased with the increased frequency
The problem of research is to identify the uses of Iraqi women for Facebook and the motives of these uses and the innovations that have been achieved from them, in view of the increasing role-played by social networking sites in the lives of individuals in general and women in particular.The research seeks to identify the following: Identify the extent to which Iraqi women use Facebook and the usage motives and innovations achieved by the interviewees When they using Facebook. This study belongs to the descriptive studies that are interested in monitoring the characteristics of a particular phenomenon to identify its characteristics and characteristics.This study adopted the survey methodology to test the hypotheses of the study and how
... Show MoreThe results of the present study showed that twenty-five samples were collected for the age group 35–40 years and four samples for the age group 65–70 years for both genders. The results showed that 48 (48%) of the samples were obtained from the hands, 16 (16%) from the legs, 12 (12%) from the abdominal area, and 10 (10%) from the chest area. The four (4%) samples were obtained from burns in the back and thighs area. The samples taken according to the cause of burns were 40 (40%) due to hot water, hot liquids, or hot steam, followed by 18 (18%) due to the use of hot tools, 15 (15%) due to fires, 12 (12%) due to electric currents, 10 (10%) due to chemicals such as strong acids, alkaline lye, paint thinner, or gasoline, and 5 (5%) due
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