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.
This work is devoted to a comparative study of the phenomenon of enantiosemy in Russian and Arabic.Everyone knows the term antonyms - words of the same part of speech, opposite in meaning, such as: Day and night, white and black, truth and lies. But in Russian, Arabic and other languages there is an interesting phenomenon, which consists in the fact that one word has two opposite meanings. Such a phenomenon in linguistics is called enantiosemy (from the Greek words enantios - "opposite" and sema - "sign")
Background: Hypothyroidism is a decrease in the production of the thyroid hormones and leads to gland dysfunction. Ashwagandha extract was used as an ayurvedic treatment and supposed to be as antihypothyroidism agent.
Objectives: to investigate the impact of ashwagandha (Ash) extract on propylthiouracil (PTU)-induced hypothyroidism in rats.
Subjects and Methods: The rats were divided into three groups, control group, PTU (hypothyroid) group (6mg/kg/day by oral route), PTU (6mg/kg/day by oral route) +Ash (50mg/kg/day by oral route) treated group. All treatment continued for
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This work aimed to design, construct and operate a new laboratory scale water filtration system. This system was used to examine the efficiency of two ceramic filter discs as a medium for water filtration. These filters were made from two different ceramic mixtures of local red clay, sawdust, and water. The filtration system was designed with two rotating interfered modules of these filters. Rotating these modules generates shear force between water and the surfaces of filter discs of the filtration modules that works to reduce thickness of layer of rejected materials on the filters surfaces. Each module consists of seven filtration units and each unit consists of two ceramic filter discs. The average measured hy
... Show MoreEl estudio se clasifica dentro los estudios teóricos sobre literatura que se ocupa del estudio de los métodos de la crítica literaria: El Estructralismo literario francés y el Formalismo ruso del siglo XX que se utilizan en la interpretación literaría. Las dos corrientes literarias estudian la literatura como ciencia que busca aplicar un método científico al estudio de la literatura. Dicho estudio trata de exponer las teorías críticas que surgen en el debate de la interpretación de los textos literarios como el de Susan Sontag, Ricoeur Paul y Mijail Bajitin, etc. Además se incluye algunos ejemplos implican el análisis estructuralista y formalista como Kafka y la tragedia Judía. y Sur Racine.
ABSTRACT:
... Show MoreThe problem of rebellion is considered one of the features of rapid changes that a society undergoes in all spheres and directions of life, especially in the realm of social relations, customs, traditions, values, and principles. Rebellion may manifest itself in rebellion against oneself, against values or traditions, or against social or governmental authority. One may find that submission plays a vital role in all of these interactions. This study deals with the problem of rebellion in the works of two renowned authors: The French Gustave Flaubert and the Israeli Amos Oz, through two main characters who share similar qualities and traits. Emma Bovary and Henna Konin demonstrate this through their rebellion against themselves, their relati
... Show MoreInnovative various Schiff bases and their Co(II), Ni(II) and Cu(II) and Hg(II) compounds made by the condensation of 4-amino antipyrine with derived aminobenzoic acid (2-aminobenzoic acid, 3-aminobenzoic acid, and 4-aminobenzoic acid ) have been prepared by conventional approaches. These complexes were described by magnetic sensibility analysis, FT-IR spectra, and molar-conductance and elemental analysis. Analytical values appeared which the mixed-ligand complexes presented ratio about 2:1 (ligand: metal) with the chelation 4 or 6. The prepared compounds offered a good effect on the organisms; bacteria Staphylococcus-aurous, Escherichia-coli and fungi C. albicans, A. niger. Also, the biological products signalize which the mixed compl
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Schiff base (methyl 6-(2- (4-hydroxyphenyl) -2- (1-phenyl ethyl ideneamino) acetamido) -3, 3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0] heptane-2-carboxylate)Co(II), Ni(II), Cu (II), Zn (II), and Hg(II)] ions were employed to make certain complexes. Metal analysis M percent, elemental chemical analysis (C.H.N.S), and other standard physico-chemical methods were used. Magnetic susceptibility, conductometric measurements, FT-IR and UV-visible Spectra were used to identified. Theoretical treatment of the generated complexes in the gas phase was performed using the (hyperchem-8.07) program for molecular mechanics and semi-empirical computations. The (PM3) approach was used to determine the heat of formation (ΔH˚f), binding energy (ΔEb
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