hrGFP Transfection into ZebraFish ZF4 cell line using Two Recombinant Proteins virD2& virE2

Document Type : Research Paper

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Abstract

Import of DNA into fish genome is generally inefficient. Therefore, one of the current challenges in fish tarnsgenesis is the development of efficient DNA delivery systems. Here we test using two bacterial proteins to target DNA to zebrafish ZF4 cell line genome. Agrobacterium tumefaciens, a soil bacterium, efficiently transfers DNA as a nucleoprotein complex to plant cells genome. Agrobacterium proteins VirD2 and VirE2 perform important functions in this process .We reconstitute this synthetic complexes consisting of the bacterial proteins VirD2, VirE2, and single-stranded DNA (ssDNA) in vitro , to test the potential of this approach to enhance gene transfer efficiency to zebrafish ZF4 cell line. Our result revealed that VirD2 and VirE2 improved the efficiency of DNA integration into zebrafish ZF4 cell line genome.

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