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A-Level Biology OCR A

Plasmid Sticky Ends Dna Complementary Membrane Allowing Explain

A plasmid can be used as a vector by cutting it with restriction enzymes, creating sticky ends complementary to the desired DNA fragment. DNA ligase then joins the two fragments, and the recombinant DNA is introduced into the host cell via transformation (using calcium chloride and heat shock) or electroporation.

I plasmidul poate fi utilizat ca vector prin tăierea acestuia cu enzime de restricție, creând capete lipicioase complementare cu cele ale fragmentului de ADN dorit. ADN ligaza apoi unește cele două fragmente, iar ADN-ul recombinant este introdus în celula gazdă prin transformare (cu clorură de calciu și șoc termic) sau electroporare.

Front Explain how a plasmid can be used as a vector.
Back Plasmids:
  • Restriction endonucleases are used to cut opne a plasmid
  • Resulting plasmid has complementary sticky ends to the sticky ends of the DNA fragment
  • Once the complementary bases of two sticky ends are lined up, DNA ligase forms phosphodiester bonds between the sugar and phosphate groups on two strands of DNA, joining them together
  • Recombinant DNA transferred into host cell via transformation:
    • Culturing bacteria in calcium-rich solution and increasing temperature causes bacterial membrane to become more permeable, allowing plasmids to enter
    • Electroporation where small electrical current is applied to bacteria, making membrane porous and allowing plasmid to move in

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