Question
How bacterial host cell is made competent to accept hydrophilic DNA, which allow DNA to pass through the cell membrane?
✓
Answer
→ Since DNA is a hydrophilic molecule, it cannot pass through cell membranes.
→ In order to force bacteria to take up the plasmid, the bacterial cells must first be made 'competent' to take up DNA.
→ This is done by treating them with a specific concentration of a divalent cation, such as calcium, which increases the efficiency with which DNA enters the bacterium through pores in its cell wall.
→ Recombinant DNA can then be forced into such cells by incubating the cells with recombinant DNA on ice, followed by placing them briefly at 42°C (heat shock).
→ Finally, putting them back on ice.
→ This enables the bacteria to take up the recombinant DNA.
→ Due to heat and cold effect the pore size of cell membrane is manipulated.
→ In order to force bacteria to take up the plasmid, the bacterial cells must first be made 'competent' to take up DNA.
→ This is done by treating them with a specific concentration of a divalent cation, such as calcium, which increases the efficiency with which DNA enters the bacterium through pores in its cell wall.
→ Recombinant DNA can then be forced into such cells by incubating the cells with recombinant DNA on ice, followed by placing them briefly at 42°C (heat shock).
→ Finally, putting them back on ice.
→ This enables the bacteria to take up the recombinant DNA.
→ Due to heat and cold effect the pore size of cell membrane is manipulated.
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