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.
16S rRNA gene sequence examination is an effective instrument for characterization of new pathogens in clinical specimens. Akey component of colonization, biofilm formation, and protection of the pragmatic human pathogen Pseudomonasaeruginosais the biosynthesis of the exopolysaccharide Psl.Extracellular polysaccharides,biofilm, are secreted by microorganisms into the neighboring environment and are significant for surface attachment and keeping structural safety within biofilms.Biofilm production is an important technique for the survival of P. aeruginosa,and its association with antimicrobial resistance represents a defy for patient therapeutics. The aim of the current research is to assess the antibiotic resistance manner and distribution
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There is an increasing interest in the use of plant extracts as therapeutic agents, particularly their capacity to inhibit the growth of pathogenic microorganisms. In this study antibacterial effect of Malva sylvestris, Anastatica hierochuntica and Vitis vinifera leaves extracts were evaluated against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Staphylococcus aureus and Proteus mirabilis. The in vitro antibacterial activity was performed using agar well diffusion method and the minimum inhibitory concentration (MIC) was determined by microtitration technique. The result indicated that the extract of V. vinifera leaves inhibited with the growth of gram-positive bacteria, as well as gram-negative bacteria while the extract
... Show MoreChemical analysis for evaluation of Nigella sativa L. (black cumin) seeds showed a composition of Fat 39% ; Protein 28% ; Carbohydrate 21% ; Moisture 6% and Ash 4.5% . It was found that the black seed contains the following mineral element : Magnesium 0.26 gm /100gm seed ; Calcium 0.25 gm /100gm seed and Iron 25 ?g / gm /100gm seed ; zinc 4.51?g /gm /100gm seed and Copper 3.60 ?g /gm /100gm seed. The analysis also showed that mineral element I. e. ; lead ; Cobalt ; Nickel ; Chrom ; Cadmium and Aresenic are not present . It was found that the fat of the black seed contains the following fatty acids : Myristic 2.8%; Palmtic 16.6%; Stearic 0.8 % ; Oleic 13.79% ; Linoleic 64.2% and Arachidic 1.9% .
Expanded use of antibiotics may increase the ability of pathogenic bacteria to develop antimicrobial resistance. Greater attention must be paid to applying more sustainable techniques for treating wastewater contaminated with antibiotics. Semiconductor photocatalytic processes have proven to be the most effective methods for the degradation of antibiotics. Thus, constructing durable and highly active photocatalytic hybrid materials for the photodegradation of antibiotic pollutants is challenging. Herein, FeTiO3/Fe-doped g-C3N4 (FTO/FCN) heterojunctions were designed with different FTO to FCN ratios by matching the energy level of semiconductors, thereby developing effective direct Z-type heterojunctions. The photodegradation behaviors of th
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Videogames are currently one of the most widespread means of digital communication and entertainment; their releases are attracting considerable interest with growing number of audience and revenues each year. Videogames are examined by a variety of disciplines and fields. Nevertheless, scholarly attention concerned with the discourse of videogames from a linguistic perspective is relatively scarce, especially from a pragma-stylistic standpoint. This book addresses this vital issue by providing a pragma-stylistic analysis of the digital discourse of two well-known action videogames (First Person Shooter Games). It explores the role of the digital discourse of action videogames in maintaining real-like interactivity between the game and the
... Show MoreBacteriocin is an important antimicrobial peptide that can be used in industrial and medical fields due to its characteristics of antibacterial, food preservation and anticancer activities. Fifty isolates of Bacillus sp were collected from different soil samples which were already recognized via morphological and biochemical identification process. The isolates were screened for bacteriocin production effective against Staphylococcus spp in order to select the highest producing isolate. The isolate NK16 showed the maximum bacteriocin production (80 AU/ml) which was further characterized as Bacillus subtilis NK 16 through using API identification system (API 20E and API 50CHB). Then, next step was to detect the optimal conditions for maximum
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Gingival crevicular fluid (GCF) may reflect the events associated with orthodontic tooth movement. Attempts have been conducted to identify biomarkers reflecting optimum orthodontic force, unwanted sequallea (i.e. root resorption) and accelerated tooth movement. The aim of the present study is to find out a standardized GCF collection, storage and total protein extraction method from apparently healthy gingival sites with orthodontics that is compatible with further high-throughput proteomics. Eighteen patients who required extractions of both maxillary first premolars were recruited in this study. These teeth were randomly assigned to either heavy (225g) or light force (25g), and their site specific GCF was collected at baseline and aft
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This study aimed to evaluate the IHC expression of CDX2 protein in HGC patients and control groups and also to study the correlation between IHC expression of the CDX2 and different clinicopathological variables such as: age, gender, histopathological subtype, grade, and stage of the tumor in HGC cases. the retrospectively sectional study for the period from 2014 to 2018 included a total of 60 formalin fixed paraffin embedded blocks of the HGC tissue (partial or total gastrectomy specimens) that collected from the archived materials of the Department of Pathology of Baghdad Teaching Hospital and the Center of Gastrointestinal and Hepatic Diseases, and also some samples were collected from other private laboratories. The IHC expression of th
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