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Bacterial Transformation
Introduction:
Purpose: The objective of this Bacterial Culture Transformation is: to observe bacterial growth under various conditions including the transformation of bacteria; to understand the process in which this transformation occurs, as well as the results and consequences of the transformation using pBlu plasmid, Ampicillin, and X-Gal Protocol.
Transformation is the "genetic alteration of a cell resulting from the uptake, genomic incorporation, and expression of a foreign genetic material (DNA)" (en.wikipedia.org/wiki/transformation, "Definition of Transformation_genetics"). Transformation in bacertial cells occur when a cell incorporates naked DNA into its genetic material; in a lab setting. This is done so by placing the mixtures of transformation solution and plasmid DNA (in pBLU tube only) on ice, then transferring them into hot water for about fifty seconds, and then back to the ice- this procedure is called heat shock and "increases the permeability of the cell membrane to DNA". This is new genetic material that is incorporated into the bacterial plasmid by the agent.
Genetic transformation occurs when DNA is taken in and expressed by a cell from a living organism. Three basic things are needed to perform genetic transformation: a host, a vector, and a method to select and isolate the transformed organisms. The host is the organism that will take in the DNA. The vector is the means of transporting the DNA into the host. A common method for selecting the transformed organisms is to introduce them to an environment containing what they are resistant to (once they transform) because only the transformed organisms will be able to survive in that environment.
In this experiment, we will perform genetic transformation of the bacteria Escherichia coli (E. coli), therefore it is properly defined as bacterial transformation. E. coli will be our host. It is a good host for transformation...