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.
Absract
The letters of the Al-Safa Brothers contain many tales, in which each story is a literary text that is independent from the other texts, but it constitutes a unified entity in its entirety, because these stories contain recurring structural elements that go into building each one of them. The stories - the subject of the analysis - were not keen on something that was keen on adhering to the Sufi vision and its apparent effect on the tales of the messages, starting with the announcement of the narration to the narrator and his attempt to persuade the recipient of the truth of what is being told and its realism, and made him believe it. The stories correspond to the formal construction of the ideal style defined by th
... Show MoreAbstract
This study is aim to :
- Put a standard to evaluate the altruistic behavior in kindergarten children.
- Know level of the altruistic behavior in the kindergarten children according to their mother "s points of view.
- Know the level of the altruistic behavior in the kindergarten children according to their teacher "s points of view.
- The differences in the level of altruistic behavior in kindergarten children between their mothers & teachers points of view.
The sample consists of (120) children of kindergarten children in Baghdad. The researcher has built a to
... Show MoreIntestinal parasites present in freshwater from the Al- Fallujah, Al- Habbaniyah and Al-Alwarar, of the Euphrates river in Iraq are Cryptosporidium spp (25.3%), Giardia sp (3.3%), Eimeria sp (3.3%), Pinworm eggs (3.3%), Naegleria sp (15.3%), Lecane niwati (1.3%), Trichomonas hominis (19.3%), Acanthamoeba spp (24.6%), Entamoeba coli (20.6%), Balantidium coli (12%), Ascaris sp (3.3%), Volvox sp (26%), Chilomastix mesnili (4%), Pelomyxa palustris (2.6%), Trinema enchelys (2.6%), Actinophrys Sol (7.3%), Amobea Vespertilio (9.3%), Rhabditea (5.3%), paramecium bursaria (9.3%), cyst of cestode (6%), Oocyst protozoa (16%), Euglena gracilis (10.6%).were isolated. The study's goal was to isolate some of the parasites that pollute the Euphrat
... Show More
(1)
In this research, Schiff bases derived from the reaction of anthrone with different heterocyclic amines have been described. The resulted Schiff base compounds were reacted with various nucleophiles in order to obtain new heterocyclic derivatives. Chemical structures of all products were confirmed by IR, 1H-, 13C-NMR spectral data and elemental analysis. All synthesized compounds were in vitro tested against a standard strain of pathogenic microorganism including Gram +ve bacteria (Staphylococcus aureus), Gram –ve bacteria (Escherichia coli), and fungi (Candida albicans).
This study aims to identify the concepts of financial crisis and its reasons of creation , also explain the effects of the accounting disclosure and the International Accounting Standards in current financial crisis, In addition to, indicate the role of accounting in the reform of the financial system from the impact of financial crisis.
The methodology of this study orientied to two main aspects, the first is an identifying approach through exploring the opinion of financial experts, the second aspect is based on an analytical approach to satisfy the requirements of experts to get there opi
... Show MoreSoils at Al-Koot-Btera were choosen to determine their sedimentary
environments. It is found that there are , five soil series and as mentioned :
MF11-MW9-DM97-DM57-DF95 . The five found soil series are of internal
well drained ,moderate and imperfect. Their textures vary in moderately,fine
and moderately fine.
indicating that sediments rang from poorly to very poorly sorting. Values of median
diameter Md Ø range between 4.11-7.80 Ø .The relation between the sorting
and median diameter shows that 95.24% of samples is a sedimentary
environment of aquite river , while 4.76% is aeolian sediments.
The values of meso to platy kurtic of most horizon materials rang
between 0.67-1.26 Ø .That is to say the samples
The present work represents description of three new species of genus Anthrenus
Geoffory from Iraq, these are : A. aradensis sp. nov., A. fabrici sp. nov. and A.
unicolor sp. nov. Locality, host plants and date of collection were given.
Six isolates of Bacillus thuringiensis were isolated from Iraqi soil characterized as non- insecticidal and non- hemolytic parasporal inclusion proteins. Bacterial isolates were propagated on nutrient broth. Then, the parasporal inclusion proteins were extracted and processed with proteinase K and trypsin. The major protein segments produced of 64KDa were characterized and tested for cytocidal activity against human leukemic T- cells (CLL) (Chronic lymphoid leukemia). Results indicated that the treated parasporal proteins of four isolates (Bt2, Bt3, Bt4 and Bt6) showed strong cytotoxicity with no significant differences between normal lymphocytes and leukemic lymphocytes. Two isolates BtA1 and BtA5 show discriminative cytotoxicity between n
... Show More
(2)
(1)
Some of the characters of the Staphylolysin A and D enzymes purified from Pseudomonas aeruginosa P16 and P5 respectively were studied, the molecular weights of Staphylolysin A and D were 20.417 kilo dalton and 23.988 kilo Dalton respectively by SDS- polyacryl amide gel electrophoresis. The optimum pH for staphylolysin A activity was found to be 8 which gives higher activity reaches 150 unit/ml, and for enzyme stability was 7.5-8.5 in which the enzyme nearly retained its full activity, while it was 9.5 for staphylolysin D that gives higher activity of 16 unit/ml,and 8.5-9.5 for enzyme stability in which the enzyme nearly retained its full activity, Maximum activity of two enzymes was obtained at 40C in which the specific activity for st
... Show More
197 vaginal swabs were collected from women of different ages. (60) Isolates of Candida albicans (30.4%) were obtained, and the other species of Candida represent (18.27%). Bacterial infections showed (41.11%), and infection with Trichomonas vaginalis was (2.03%). Ten isolates of C. albicans were chosen randomly for farther study which include two virulence factors tendency of adhesion wich showed a percentage of (52%) to(32%) , and the ability to produce phospholipaze enzyme and it’s activity which showed (50%) of the isolates have the ability to produce the enzyme in different degrees .