Concept explainers
Describe how the process of gene doning results in a cell clone contaning a recombinant plasmid.
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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- Assume that there are horizontal gene transfers between two completely different bacterial species. In one case it is a plasmid that is transmitted via conjugation, in the other case it is it is a part of the bacterial chromosome that is transferred via transformation. In which of the two cases is it likely that the transferred DNA will be present? left and can function in the recipient cells? Explain the biological background to your answerarrow_forwardIn order to construct a genome library, which statement most accurately describes the correct order of steps in the process? Fragment chromosomal DNA, digest plasmid vector, ligate genomic DNA and digested plasmid vector, transform ligation into bacteria, select for plasmid marker Fragment chromosomal DNA and plasmid vector, ligate genomic DNA and plasmid vector, transform ligation into bacteria, select for gene marker Fragment chromosomal DNA and plasmid vector, ligate genomic DNA and plasmid vector, transform ligation into bacteria, select for plasmid marker Fragment chromosomal DNA, ligate genomic DNA and plasmid vector, transform ligation into bacteria, select for plasmid markerarrow_forwardPlasmids are the only vectors currently available for use in recombinant procedures.True or false?arrow_forward
- explain how Plasmids Carry Genes in Addition toThose in the Bacterial Chromosomearrow_forwardExplain the basis for the following statement: Transcription of two genes on a plasmid can occur without the concomitant action of a topoisomerase, but only if those two genes are oriented in opposite directions.arrow_forwardIn bacterial transformation, the purpose of having antibiotic within an agar plate is to: Select one: confirm which plasmids been have successfully ligated with a gene of interest. isolate bacteria which have been successfully transformed with the plasmid. indicate which plasmids were successfully digested by the endonuclease. act as a substrate which will be cleaved and produce a blue product when ligation is unsuccessful. show which plasmids contain the lacZ gene.arrow_forward
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