Question

while conducting the transformation in bacteria, why doesn't the recipient or the F- cell consider the DNA or plasmid as a foreign particle and degrade it ?
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Answer 1

Answer:

The naked plasmids can be attacked by endonucleases in the cells. So, when doing PEG-mediated protoplast transformation or yeast transformation, 'carrier DNA' (ex. Salmon Sperm DNA) are usually added with your plasmids to transfect cells. One of the reasons to add carrier DNA is to reduce the risk of plasmid-of-interest degradation inside the cells.

2. For Agrobacterium transformation, the T-DNA is transferred as a Protein-TDNA complex (or T-complex). From research results, the protein molecules seems to provide the protection for the single-stranded T-DNA. See the attached article (yellow highlight): "VirE2 association with ssDNA molecules in vitro was shown to protect them from exonucleolytic degradation".

Answer 2

1. The naked plasmids can be attacked by endonucleases in the cells. So, when doing PEG-mediated protoplast transformation or yeast transformation, 'carrier DNA' (ex. Salmon Sperm DNA) are usually added with your plasmids to transfect cells. One of the reasons to add carrier DNA is to reduce the risk of plasmid-of-interest degradation inside the cells.

2. For Agrobacterium transformation, the T-DNA is transferred as a Protein-TDNA complex (or T-complex). From research results, the protein molecules seems to provide the protection for the single-stranded T-DNA. See the attached article (yellow highlight): "VirE2 association with ssDNA molecules in vitro was shown to protect them from exonucleolytic degradation".

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