Concept explainers
Describe how the process of gene doning results in a cell clone contaning a recombinant plasmid.
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To explain: The gene cloning process that produces a cell clone having a recombinant plasmid.
Concept introduction:
To study a particular protein, multiple copies of the gene fragment encoding that protein are synthesized. These gene fragments are then inserted into the expression vector to synthesize a large quantity of protein that is encoded by the gene. This is known as gene cloning.
Explanation of Solution
For cloning a eukaryotic gene, the bacterial plasmid vector and the gene segment to be cloned are cleaved with the same restriction sites. Fig. 1 shows the cloning procedure using EcoRI restriction enzyme as both the sequences have their restriction sites. The arrows between the bases in Fig. 1 represent the restriction sites. The restriction digestion of the linear gene sequence and the plasmid sequence produces sticky ends. The digested gene sequence is ligated into bacterial plasmid and the recombinant plasmid is synthesized. This recombinant plasmid is reintroduced into the host cells (bacteria), where the eukaryotic gene expresses and forms proteins. The bacterial cells containing the recombinant plasmids will divide to form cell clones.
Pictorial representation:
Fig. 1 shows the cloning procedure using a gene sequence and bacterial plasmid.
Fig. 1 Cloning procedure
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