Question

1). Explain DNA transformation?

2). In which of the 3 plates namely LB, LB/AMP, LB/AMP/Ara was DNA transformed?

a). Which plates are the control plates? Why do we need the control plates?

3). Only one plate has transformants. This is the experimental plate. Which one is it? Why do only transformants grow on this plate?

Answer 1

DNA transformation is a process of horizontal gene transfer by which some bacteria take up foreign genetic material from the environment.

Plasmid or vector DNA transformation implies uptake of DNA into bacterial, yeast or plant cells, while transfection is a term usually reserved for mammalian cells. Typically the method for transformation of a DNA construct into a host cell is chemical transformation, cell are made competent by treatment with divalent cations such as calcium chloride, which make the bacterial cell wall more permeable to DNA . Heat shock is used to temporarily from pores in the cell membrane, allowing transfer of the exogenous DNA into the cell. In electroporation , a short electrical pulse is used to make the bacterial cell temporarily permeable. Particle bombardment , is typically used for the transformation of plant cells. Gold or tungsten particles are coated with the DNA construct and physically forced into the cell by gene gun.

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