Biology Module 2 - 4.aspects of genetic engineering 4.2 explain the steps involved in recombinant DNA technology

4.2 explain the steps involved in recombinant DNA technology

• the plasmid is isolated and taken from the bacterium

• DNA making up the plasmid is cut using restriction enzyme

• Insulin gene is isolated and cut using same restriction enzyme

• length of insulin gene is then inserted into the vector in this case the plasmid is the vector and they complementary base pair because of sticky ends which are short unpaired bases at each end of DNA

• recombinant DNA is then treated with the enzyme ligase which is used to link sticky ends

• the host bacterium is treated with calcium ions, then cooled and given a heat shock. This allows the plasmid to pass through the bacteria cell membrane

• recombinant plasmid is then added to bacterium and the plasmids then replicate

• the bacteria then divides by binary fission so that each daughter cell has several copies of the plasmid