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
Abstract
This paper concerned with study the effect of a graphite micro powder mixed in the kerosene dielectric fluid during powder mixing electric discharge machining (PMEDM) of high carbon high chromium AISI D2 steel. The type of electrode (copper and graphite), the pulse current and the pulse-on time and mixing powder in kerosene dielectric fluid are taken as the process main input parameters. The material removal rate MRR, the tool wear ratio TWR and the work piece surface roughness (SR) are taken as output parameters to measure the process performance. The experiments are planned using response surface methodology (RSM) design procedure. Empirical models are developed for MRR, TWR and SR, using the analysis
... Show MoreRemoval of Congo red, Rhodamine B, and Dispers Blue dyes from water solution have been achieved using Flint Clay as an adsorbent. The adsorption was studied as a function of contact time, adsorbent dose, pH, and temperature under batch adsorption technique. The equilibrium data fit with Langmuir, Freundlich and Toth models of adsorption and the linear regression coefficient R2 was used to elucidate the best fitting isotherm model. Different thermodynamic parameters, namely Gibb’s free energy, enthalpy and entropy of the on-going adsorption process have also been evaluated. Batch technique has been employed for the kinetic measurements and the adsorption of the three dyes follows a second order rate kinetics. The kinetic investigations al
... Show MoreThe current study aims to identify the level of adaptation of the curriculum for visually handicapped pupils from the point of view of their teachers as well as to identify the level of adaptation of the educational programs offered to visually handicapped pupils from the point of view of their teachers. In order to achieve the goals of the research, the researcher developed two tools to measure the adaptation of curricula and educational programs for pupils with visual handicapped from the point of view of teachers. The sample included (60) teachers from Al Noor Institute for Blinders in Baghdad, the two tools were applied to them, and then the Reliability and Validity coefficients were extracted. The results sh
... Show MoreThe current study was conductedas a pot experiment to determine the effect of soil texture on biological nitrogen fixation (BNF) of six most efficient local isolates, specified, of Bradyrhizobium. Cowpea (Vignaunguiculata L.), as a legume host crop, was used as a host crop and 15N dilution analysis was used for accurate determination of the amount of N biologically fixed under experimental parameters specified. Soils used are clay loam, sandy clay loam and sandy loam. Biological Nitrogen Fixation (BNF), in different soil textural classes, was as in the following order: medium texture soil > heavy texture soil > light textured soil. Statistical analysis showed that there is a significant variation in BNF % among six Iraqi isolates in the th
... Show MoreIn the 1970s, the world knew the long-tailed nesokia Nesokia bunnii (Khajuria, 1981) (Rodentia, Muridae) from the Mesopotamian marshes of Garden of Eden in Southern Iraq. This distinct rodent was known from only five voucher specimens collected at the confluence of Tigris and Euphrates Rivers in southern Iraq while its occurrence in Southwestern Iran had
never been reported. In the 1990s, a large extent of its natural habitat was catastrophically desiccated and the animal was last seen in the 1970s. Since then, the status of this elusive rodent was shrouded in mystery. In 2007, an extraordinary photograph of a carcass of this species came to the light from Hawizeh Marsh which was interpreted as concrete evidence of the species’ pers
Coupling reaction of 4-amino antipyrene with 4-amino benzoic acid gave bidentate azo ligand. The prepared ligand was identified by Microelemental Analysis, 1HNMR, FT-IR and UV-Vis spectroscopic techniques. Treatment of the prepared ligand with the following metal ions (CoII, NiII, CuII and ZnII) in aqueous ethanol with a 1:2 M:L ratio and at optimum pH, yielded a series of neutral complexes of the general formula [M(L)2]Cl2 . The prepared complexes were characterized using flame atomic absorption, (C.H.N) Analysis, FT-IR and UV-Vis spectroscopic methods as well as magnetic susceptibility and conductivity measurements. Chloride ion content was also evaluated by (Mohr method). The nature of the complexes formed were studied following the mol
... Show MoreHighly-fluorescent Carbon Quantum Dots (CQDs) are synthesized in simple step by hydrothermal carbonization method of natural precursor such as orange juice as a carbon source. Hydrothermal method for synthesized CQDs requires simple and inexpensive equipment and raw materials, thus this method are now common synthesis method. The prepared CQDs have ultrafine size up to few nanometers and several features such as high solubility in water, low toxicity, high biocompatibility, photo-bleaching resistant, Chemical inertness and ease of functionalization which qualifies it for use in many applications such as bio-imaging, photo-labeling and photo-catalysis.
This research demonstrates the
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