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.
Calendula officinalis L. (Asteraceae) known as marigold is known to have several pharmacological activities and used for the treatment of several diseases as measles, jaundice, constipation and several inflammations. Marigold flowers contain several chemical constituents mainly flavonoids, triterpenoids and essential oil. In this study marigold flowers cultivated in Iraq had been investigated for its flavonoids content. The study revealed the presence of quercetin and kaempferol glycosides and the absence of myricetin glycosides. The flowers were extracted with ethanol 70% fractionated with different solvent and the flavonoids were isolated by preparative HPLC. The isolated flavonoids were identified by measuring melting points, UV, IR,
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Ticks (Acari: Ixodidae) are ectoparasites that infest livestock in every geographic region of the world and are vectors of several viral, bacterial, and protozoan pathogens to both animals and humans. There is little information is available is about tick presence in Buffalo Bubalus bubalis (Linnaeus, 1758) (Artiodactyla, Bovidae) in Iraq. The current study determined the species of ticks parasitizing Buffalo in some central and southern regions included: Baghdad (Al Fathelia), Karbala (Al-Hussainia), Wasit (Kut and Al-Suwairah), Al-Qadisia (Al- Diwaniyah, Al- Saniya, Al-Mihnawea, and Afak), Thi Qar (Al-Nasiriyah and Al-chibayish), Missan (Amara and Qalaat Salih) and Basrah (Al-Haretha, Al-Madena and Al-Deer). A total of 150 Buffal
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The -multiple mixing ratios of γ-transitions from levels of populated in the are calculated in the present work by using the a2-ratio methods. We used the experimental coefficient (a2) for two γ -transitions from the same initial state, the statistical tensor, which is related to the a2-coefficient would be the same for the two transitions. This method was used in a previous work for pure transitions or which can be considered pure. In these cases the multiple mixing ratios for the second transition ( ) equal zero, but in our work we applied this method for mixed γ-transitions and then the multiple mixing ratio ( ) is known for one transition. Then we calculate the ( ) value and versareversa. The we
... Show MoreThe root lesion nematode, Pratylenchus neglectus, is one of the most damaging nematodes to affect wheat worldwide. The nematode is widely distributed in Montana, primarily affecting winter wheat within the state. Managing the nematode primarily involves rotation to resistant and moderately resistant crops (peas, lentils, and barley). A nematode survey was conducted across the state nearly 10 years after an initial survey, to reassess the nematode threat and assess the impact of changing trends in crop rotations. To assess the broad applicability of rotation crops to control P. neglectus across Montana, greenhouse trials were conducted to challenge rotational crops using eight populations of P. neglectus collected from geographicall
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Due to the deliberate disposal of industrial waste, a great amount of petroleum hydrocarbons pollute the soil and aquatic environments. Bioremediation that depends on the microorganisms in the removal of pollutants is more efficient and cost-effective technology. In this study, five rhizobacteria were isolated from Phragmites australis roots and exposed to real wastewater from Al-Daura refinery with 70 mg/L total petroleum hydrocarbons (TPH) concentration. The five selected rhizobacteria were examined in a biodegradation test for seven days to remove TPH. The results showed that 80% TPH degradation as the maximum value by Sphingomonas Paucimobilis as identified with Vitek® 2 Compact (France).
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In this paper flotation method experiments were performed to investigate the removal of lead and zinc. Various parameters such as pH, air flow rate, collector concentrations, collector type and initial metal concentrations were tested in a bubble column of 6 cm inside diameter. High recoveries of the two metals have been obtained by applying the foam flotation process, and at relatively short time 45 minutes . The results show that the best removal of lead about 95% was achieved at pH value of 8 and the best removal of zinc about 93% was achieved
at pH value of 10 by using 100 mg/l of Sodium dodecylsulfate (SDS) as a collector and 1% ethanol as a frother. The results show that the removal efficiency increased with increasing initial m
The study was performed to isolate and identify the Myxococcus
xanthus from (50) samples of grave soils .Special growth conditions had been used to support the growth of M.
xanthus and to suppressed the growth of other microorganisms like (Drying , High concentration of antibiotics and specific growth media)
M. . xanthus isolates had been subjected to the morphological, cultural and biochemical examinations for identification . Results obtaind could be summarized as follows : 1. Myxobacteria were found as normal flora inhabitants of the arid soils. 2. Ten local isplates of M. xanthus out of (50) soil samples were isolated
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Feasibility of biosorbent of England bamboo plant origin was tested for removal of priority metal ions such as Cu and Zn from aqueous solutions in single metal state. Batch single metal state experiments were performed to determine the effect of dosage (0.5, 1 and 1.5 g), pH (3, 4, 4.5, 5 and 6), mixing speed (90, 111, 131, 156 and 170 rpm), temperature (20, 25, 30 and 35 °C) and metal ion concentration (10, 50, 70, 90 and 100 mg/L) on the ability of dried biomass to remove metal from solutions which were investigated. Dried powder of bamboo removed (for single metal state) about 74 % Cu and 69% Zn and maximum uptake of Cu and Zn was 7.39 mg/g and 6.96 mg/g respectively, from 100 mg/L of synthetic metal solution in 120 min. of contact t
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