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
Receipt date:11/19/2020 acceptance date:10/1/2021 Publishing date:12/31/2021
This work is licensed under a Creative Commons Attribution 4.0 International License.
Human rights are one of the basic concepts of modern and contemporary times, and political thought as the origin of political science and its source of knowledge have impact in this science and its branches
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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
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The aim of the research is to highlight the role of electronic human resources management practices in the sustainability of knowledge capital as one of its success factors, as well as the diagnosis and interpretation of the relationship between research variables and their dimensions. The research problem is that the University of Babil implements some electronic human resources management practices not in a complete way, The level of its application and the problems it faces, as well as the extent to which these practices reflect the knowledge capital and sustainability in the university, and highlights the importance of research as it is concerned with the electronic aspects and achieve the competitive advant
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This is a case–control study involved 50 MM patients attending Hematology Center, Bag
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This study aimed to investigate Human parvovirus B19 (PVB19) prevalence in β-TM patients by serological and molecular means.
This is a cross-section