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Transcript
Biotechnology • various tools in biotechnology have made it possible for humans to manipulate and control DNA sequences. • recombinant DNA is a fragment of DNA which contains sequences that come from at least 2 different sources. (ie. A gene for human insulin located in a bacterial DNA). CSI Restriction Enzymes (Endonucleases) • Enzymes that are able to cut double stranded DNA at specific sequences. • They originate from bacteria and are used in their native environment to destroy (by chopping up) any DNA that is not property of the bacteria. • Restriction enzymes will cut DNA at a specific sequence (called a recognition site). • One example, EcoRI, cuts DNA at the following sequence. • Most recognition sites (as above) are 4-8 base pairs long and are palindromic (read the same bases 5’-3’ on one strand as they do 5’-3’ on the other strand). • EcoRI will scan along a DNA strand until it finds it’s recognition site and then it breaks the phosphodiester bond between G and A, resulting in the formation of two DNA strands instead of one. Creepy Restriction Enzyme Movie • some restriction enzymes (like EcoRI) produce cuts in the DNA that result in the formation of sticky ends on the DNA fragments that are formed. • sticky ends indicates that unpaired bases are left hanging off the cut. other restriction enzymes produce blunt ends, that is, the DNA is cut directly down the middle. • cuts that produce sticky ends are more preferable to a biotechnologist because they can be joined to other sticky end fragments with the help of H-bonding between complimentary bases. Plasmid Mapping Using restriction enzymes – it is possible to create a map of a plasmid that will enable the insertion of a known gene at a particular location – this is called molecular cloning.
