List and discuss the steps in the genetic engineering, isolation, and cloning of a target gene. Include the characteristics and experimental role of bacterial plasmids.
Genetic engineering is a process of manufactering bacteria plasmids using DNA sequences that are design for a particular purpose. The terms used to describe the process of creating new plasmids is called plasmid recombination. Bacterial plasmids are circular DNA and are very simplestic. This the reasons why they are used.
In order to design an efficent plasmid you will need to have:
1. Origin of replication- the cell machinery needs to know where to start the transcription process
2. Target gene- this gene tells the vector or plasmid where to target. This is useful in drug delivery models
3. Reporter gene- this gene tells us where the gene is what kinds are proteins are being produced. Examples of reporter gene is GFP (or green flourescent protein)
4. A gene of interest- this the gene that you want to test the effects of. This gene is designed and inserted into the plasmid.
There are many steps in making a recombinant plasmid:
1. Start with an pre-designed vector that is premade
2. Use restriction enzymes that cut the plasmid in the correct places so that you can insert your desired fragments into the places. Restriction enzymes act like scissors.
3. The fragments are ligated (or pasted) into the plasmid one by one. Depending on the type of vector, it can take months to add all the pieces into the vector.
4. Complete package contains all of the neccessary parts.