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.
Introduction and Aim: Cancers are a complex group of genetic illnesses that develop through multistep, mutagenic processes which can invade or spread throughout the body. Recent advances in cancer treatment involve oncolytic viruses to infect and destroy cancer cells. The Newcastle disease virus (NDV), an oncolytic virus has shown to have anti-cancer effects either directly by lysing cancer cells or indirectly by activating the immune system. The green fluorescent protein (GFP) has been widely used in studying the anti-tumor activity of oncolytic viruses. This study aimed to study the anticancer effect of a recombinant rNDV-GFP clone on NCI-H727 lung carcinoma cell line in vitro. Materials and Methods: The GFP gene was inserted t
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Most countries in the world particularly developing countries, including Iraq, facing extremely dangerous problem with social and political dimensions, which is the emergence of the food crisis problem ,the decrease in domestic food production in Iraq isn't meet the needs of its population food, due to the fact that the agricultural sector suffers from multiple natural ,economic and human problems .It is still below the level required to meet the needs of the population of food ,since food at the forefront of priorities needed by the human . This represents indispensable basic necessity , so the responsibility of its availability permanently in appropriate&nb
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The issues of journalists and media employees in general and photojournalists in particular have become important issues, especially as those issues are closely linked to the success or failure of the media process.
This research deals with (the issues of Iraqi photojournalists working in local and foreign institutions in Iraq - a case study in 2012), because of the ambiguity in identifying those issues, which focused on the issues of this research.
This was done through the research community of members of the Association of Iraqi photojournalists in Baghdad exclusively of (64) photographers and television photographers to identify the problems encountered in their work
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This study was aimed to monitor oral zinc sulfate role on local cervical proinflammatory cytokines in HPV-infected women comparing with these cytokines before treatment application. A cervical secretion got from 28 infected women before and after treatment with zinc sulfate, these samples assessed various markers of inflammation including interleukin-1β, IL-8, and IL-12. Results manifested that improve and clear the cervical HPV infections after three months of zinc treatment 46.43% and 21.43%, respectively. Viral infections with single and multiple HPV high-risk types are raising of studied cytokines after 3-month compared with single HPV low-risk type. Moreover, this increasing was statistically significant only in IL-12 and IL-1. Women
... Show MoreIn light of increasing demand for energy consumption due to life complexity and its requirements, which reflected on architecture in type and size, Environmental challenges have emerged in the need to reduce emissions and power consumption within the construction sector. Which urged designers to improve the environmental performance of buildings by adopting new design approaches, Invest digital technology to facilitate design decision-making, in short time, effort and cost. Which doesn’t stop at the limits of acceptable efficiency, but extends to the level of (the highest performance), which doesn’t provide by traditional approaches that adopted by researchers and local institutions in their studies and architectural practices, limit
... Show MoreBackground: Postoperative pain is one of the main complications following impacted mandibular third molar (IMTM) surgery. Objectives: The aim of this study was to assess the effect of the local application of bupivacaine on reducing early postoperative pain following IMTM surgery. Material and methods: A prospective, single-blinded, randomized controlled study was conducted on 40 patients who had undergone the surgical removal of an IMTM under local anesthesia. In the study group (n = 20), absorbable gelatin sponge (AGS) soaked in 3 mL of 0.5% plain bupivacaine hydrochloride was locally applied in the post-extraction socket. In the control group (n = 20), AGS soaked in 3 mL of normal saline was used. Pain intensity was assessed using a pa
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The survey showed sample opinions of officials in the general company for vegetable oils industry, and the statement of the order of the effect of these dimensions depending on the degree of importance, the questionnaire was used as a key tool in collecting data and information of the sample consisted of 30 officials, arithmetic mean, standard deviation, percentages, Spearman correlation coefficient and the global statistical analysis as a statistical methods that based on statistical program (SPSS), The researcher came to several conclusions, most important that there is high agreement by the respondents of the importance of the dimensions of building a mental picture of the company and to attract the consumer's attention, Also research
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