Biotechnology - Biotechnology Restriction Endonucleases...

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Biotechnology Restriction Endonucleases: breaks the DNA in the middle; specified breaks. Recognize specific base sequences of DNA o once sequence is found, DNA is broken at that point Host cell’s DNA protection o Methylation occurs in non-hydrogen bonding region of the nitrogenous bases occurs immediately after replication Three types of Restriction Endonucleases (Groups based upon: types of sequences) Type I: recognizes a specific sequence and then cleaves the DNA at a non- specific site (>1000 bp) away from that sequence Type III: recognizes a base sequence and cleaves the Type II: recognizes a specific sequence of DNA and cuts it within that sequence o since the cuts are always at the same place, this enzyme is useful o sites have two-fold symmetry sequences are normally palindromic Restriction Fragments o blunt ends o staggered cuts-“sticky ends” two molecules of DNA cut with the same restriction enzyme will come together by mutual attraction of their sticky ends.
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