Molecular farming has become one of the most significant implementations of modern biotechnology to generate modified plant crops to produce medicinal proteins. Agrobacterium is one plant genetic engineering tool that integrates genes of interest inside a host plant. In recent years, the need to produce recombinant proteins as therapeutics has growing rapidly, and human glucocerebrosidase is one of the proteins that is need to treat disease. In this study, specific primers were designed to amplify Hu-GBA1 gene from constructed pGEM-GBA plasmid which was cloned into the plant expression vector pCAMBIA1304. The generated recombinant pCAMBIA1304-GBA plasmid was used to transform A. tumefaciens LBA4404 and applied for transformation of sunflower cotyledon explants. Colony PCR technique was used to confirm the presence of Hu-GBA1 gene in transformed A. tumefaciens. Agrobacterium containing pCAMBIA1304-GBA was suspended in Infection Medium (IM) supplement with 200 mM acetosyringone. A bacterial suspension was used to transform sunflower cotyledons. After infection, cotyledons were co-cultivated in Co-cultivation medium (CCM), supplied with 200 mM acetosyringone without antibiotics. The cotyledons were then transferred to selection media containing 7.5 mg/L Hygromycin and 250 mg/L Cefotaxime and grown for additional 14 days at 25℃ in photoperiod of 16h L/8h D. The transformed sunflower cotyledons were successfully generated complete plant with used 6-Benzylaminopurine and Naphthalene acetic acid as growth hormones. The presence of the Hu-GBA1 gene in the genomic DNA of transgenic sunflower plant was proven by PCR as a band of 1561bp size. The GBA mRNA expression in modified sunflowers was detected by qRT-PCR compared with control GBA mRNA. Enzyme Linked Immunoassay was done on crude recombinant protein that extracted from transformed sunflower using Human Glucosylceramide ELISA Kit, the Elisa test results confirmed the production of recombinant glucocerebrosidase and the concentration of crude recombinant enzyme extracted from transformed sunflower with GBA1 gene was 0.45 ng/µl
The study of the concepts of "democracy" and "human rights" and their impact on development, has taken a prominent place in the context of general political and cultural concerns, or in specialized studies even though researchers and scholars do not agree on defining the concepts: Democracy, Human Rights, Development or deriving a global definition hindering them, and that because of the importance of this topic, which proceeds from a number of considerations standing in the forefront, of which:
First: What the world witnesses today in the first decade of this century of consecration of the unipolar theory, trying to give its concepts and perceptions through the law ((excessive force)) to the power of international
... Show MoreBy definition, the detection of protein complexes that form protein-protein interaction networks (PPINs) is an NP-hard problem. Evolutionary algorithms (EAs), as global search methods, are proven in the literature to be more successful than greedy methods in detecting protein complexes. However, the design of most of these EA-based approaches relies on the topological information of the proteins in the PPIN. Biological information, as a key resource for molecular profiles, on the other hand, acquired a little interest in the design of the components in these EA-based methods. The main aim of this paper is to redesign two operators in the EA based on the functional domain rather than the graph topological domain. The perturb
... Show MoreThe purpose of this study was to determine the impact of added soy protein humidifier concentration of different ratios of bovine Alberkr mix that was to bring the old chicken shop pure beef in five ratios
Background: syndrome X or metabolic syndrome is a collection of multiple diseases mainly visceral obesity , hypertriglyceridemia , decrease HDL level, hypertension and elevated fasting blood glucose that lead to accelerated atherosclerosis through multiple mechanisms, one of the most important is increase inflammation of the vessels manifested by elevated high sensitivity C reactive protein (hs-CRP).Objective: The aim of the study was to assess the prevalence of elevatedhs CRP in people with metabolic syndrome and atherosclerosis complication (IHD, Cerebrovascular disease, peripheral vascular disease) and metabolic syndrome without these complication.Patients and methods:;This is a cross sectional study carried out in Diabetic referral c
... Show MoreSynthesis of 2-mercaptobenzothiazole (A1) is performed from the reaction of o-aminothiophenol and carbon disulfide CS2 in ethanol under basic condition. Compound (A1) is reacted with chloro acetyl chloride to give compound (A2). Hydrazide acid compound (A3) is obtained from the reaction of compound (A2) with hydrazine hydrate in ethanol under reflux in the presence of glacial acetic acid .The reaction of hydrazide acid compound (A3) with ethyl acetoacetate gives pyrazole compound (A4). The new hydrazone compound (A5) was prepared from the reaction of compound (A3) with benzaldehyde. Reaction of compound
... Show MoreResearchers have recently increased their focus on the link between autoimmune diseases and infections. Most of the recent research indicates that silent human cytomegalovirus (HCMV), may have diverse roles in the initiation, development, and exacerbation of autoimmune diseases, such as coeliac Disease (CD) and inflammatory bowel disease. The aim of this study is to evaluate the role of HCMV infection in Iraqi patients with CD. Serum samples were obtained from 60 patients with CD, and from 60 healthy subjects. Enzyme-linked immunosorbent assay was used to determine the Anti-Transglutaminase IgG/IgA, Anti-gliadin IgA/ IgG, as well as the HCMV IgM/ IgG levels in the serum samples. Significantly higher percentage of positivity for seru
... Show MoreGingival 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
... Show MoreThe present study was undertaken in order to investigate the role of gentamicin in the gene expression of toxA in Pseudomonas aeruginosa isolated from cow mastitis. A total of ten P. aeruginosa strains originally isolated from cows infected with mastitis. Agar dilution methodology was performed to determine the minimal inhibitory concentration of gentamicin, all of which developed resistance toward gentamicin. The findings presented here demonstrated that all these strains harboured toxA depending on PCR-based assay. Nonetheless, RT-PCR technique revealed a wide variation in expression of toxA. Moreover, the cultivation of P. aeruginosa in the presence of gentamicin, significantly (P< 0.05), induced the expression of toxA, in addition to th
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