Designing E1 Deleted Adenoviral Vector by Homologous Recombination
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Abstract:
Adenoviruses are used extensively to deliver genes into mammalian cells, particularly where there is a requirement for high-level expression of transgene products in cultured cells, or for use as recombinant viral vaccines or in gene therapy. In spite of their usefulness, the construction of adenoviral vectors (AdV) is a cumbersome and lengthy process that is not readily amenable to the generation of large collection of clones. Methods: In this project, to delete E1 gene in adenovirus, an adenoviral plasmid containing lateral sites of E1 region of adenovirus was made and recombination in the 293A cells between the homologous region of this linearized plasmid and the adenovirus genome resulted in the formation of the complete adenoviral recombinant. Results: This recombination resulted in loss of E1 region and we constructed a recombinant adenovirus type 5 vector that E1 gene was deleted by homologous recombination. Conclusion: Homologous recombination is more easy and fast technique in the production of AdV.
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designing e1 deleted adenoviral vector by homologous recombination
adenoviruses are used extensively to deliver genes into mammalian cells, particularly where there is a requirement for high-level expression of transgene products in cultured cells, or for use as recombinant viral vaccines or in gene therapy. in spite of their usefulness, the construction of adenoviral vectors (adv) is a cumbersome and lengthy process that is not readily amenable to the generat...
full textDesigning E1 deleted adenoviral vector by homologous recombination.
BACKGROUND Adenoviruses are used extensively to deliver genes into mammalian cells, particularly where there is a requirement for high-level expression of transgene products in cultured cells, or for use as recombinant viral vaccines or in gene therapy. In spite of their usefulness, the construction of adenoviral vectors (AdV) is a cumbersome and lengthy process that is not readily amenable to ...
full textA simple method for constructing E1- and E1/E4-deleted recombinant adenoviral vectors.
We previously developed a two-plasmid in vitro ligation method that did not require a recombination step to produce new recombinant E1- or E1/E3-deleted adenoviral vectors. In this study, we have modified the system to improve the simplicity of vector construction and, in addition, to allow for production of an E1/E4-deleted vector.
full textNeonatal Mice Product-Specific Antibody Responses in Vaccine Carriers Induce Transgene Mucosally Delivered E1-Deleted Adenoviral
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Homologous recombination
DNA damage, in the form of DNA double-strand breaks, poses a considerable threat to genomic integrity and cell survival. If left unrepaired, a single double-strand break is sufficient to cause cell death and, if repaired inappropriately, a double-strand break may give rise to a potentially oncogenic translocation. Double-strand breaks in genomic DNA may arise accidentally in a number of ways, i...
full textRecombination: Homologous recombination branches out
Homologous recombination can be divided into three key steps: strand exchange, branch migration and resolution. The identification of a protein complex that catalyses branch migration and Holliday junction resolution argues that the mechanism of homologous recombination is conserved from bacteria to man.
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Journal title
volume 11 issue 3
pages 199- 202
publication date 2007-07
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