[17] Transformation and Transfection Flashcards
What is transformation in genetics?
The process of introducing foreign DNA into a cell.
What organisms commonly undergo transformation?
Typically bacteria and yeast.
What is transfection?
The process of introducing foreign DNA into eukaryotic cells.
What are the differences between transformation and transfection?
Transformation is used for prokaryotes while transfection is used for eukaryotic cells.
What are common methods of transformation in bacteria?
Heat shock and electroporation.
What is heat shock in the context of bacterial transformation?
A method where bacterial cells are exposed to a high temperature for a short period of time to make them permeable to plasmid DNA.
What is electroporation in the context of transformation?
A method where an electric field is applied to cells to increase permeability of the cell membrane, allowing DNA to be introduced into the cell.
What are common methods of transfection in eukaryotic cells?
Lipofection, electroporation, and viral transduction.
What is lipofection in the context of transfection?
A method where DNA is encapsulated in liposomes or lipid bubbles, which then fuse with the cell membrane to deliver the DNA into the cell.
What is viral transduction in the context of transfection?
A method where a virus is used as a vector to deliver the DNA into the cell.
What is the purpose of transformation and transfection in genetic engineering?
They are used to introduce foreign DNA into cells for the purpose of gene cloning, protein production, or to alter the cell’s characteristics.
What is a transformation efficiency?
The number of cells that take up plasmid DNA and express it, per unit of DNA used in a transformation experiment.
How is transformation efficiency important in genetic experiments?
High efficiency is desirable as it increases the chances of obtaining desired clones or cells expressing the desired gene.
What is a selectable marker in the context of transformation and transfection?
A gene introduced with the DNA of interest that provides a way to identify cells that have taken up the foreign DNA.
What is a common selectable marker used in transformation and transfection?
Antibiotic resistance genes.
How do antibiotic resistance genes function as selectable markers?
Cells that have taken up the foreign DNA can survive and grow in the presence of the antibiotic, while cells that have not will die.
What is stable transfection?
A long-term method where the foreign DNA is integrated into the host cell’s genome.
What is transient transfection?
A short-term method where the foreign DNA remains in the cell’s cytoplasm and is lost after cell division.
What are the advantages of stable transfection?
It allows for long-term expression of the foreign DNA and can be passed on to daughter cells.
How does a binary vector system work in plant genetic engineering?
The first plasmid, which contains the gene of interest, is introduced into Agrobacterium. This bacterium then transfers the gene of interest into the plant cell.
