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Q.How can we make a host cell competent to receive a foreign gene or DNA ?
Kerala DhseKerala DHSE Plus Two Board 2020Subjective· 2mImportance★★★★★
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Start your 14-day free trial to unlock the full solution →Bacterial cells are made 'competent' to take up foreign DNA by treating them with a specific concentration of a divalent cation, then briefly heat-shocking them in the presence of recombinant DNA — the CaCl2/heat-shock transformation method.
DNA is a hydrophilic molecule and cannot easily pass through the hydrophobic cell membrane of a bacterium on its own, so the host cell must first be made 'competent' to take up foreign DNA.
- Bacterial cells are treated with a specific concentration of a divalent cation, such as calcium chloride (CaCl2). This treatment increases the efficiency of DNA entry through pores created in the cell wall.
- Recombinant DNA (the plasmid/vector carrying the gene of interest) is then incubated with these competent cells on ice.
- The cells are given a brief heat shock, typically by placing them at 42°C for about 90 seconds, and then rapidly put back on ice. This heat shock causes the DNA to enter the cell through the enlarged pores. …
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