Restriction Enzymes

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Restriction Enzymes

• By Idongesit Bullard, Peter Gajda, and Serge
  Farinas

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Restriction Enzymes

• Found in bacteria and archaea, thought to be
  evolutionary defense mechanism
• Recognize particular nucleotide sequence and cuts
  double stranded DNA at this point
• Extremely important for molecular biology
  cloning and restriction site mapping

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Blunt vs Sticky Ends

• Blunt and sticky ends are produced by the
  restriction enzyme it depends on the enzyme.
• Blunt ends are produced when the enzyme makes a
  clean cut at the recognition site.
• --------------
  
  ACCGATTAGCGAC ACCGATTA
  GCGAC TGGCTAATCGCTG
  TGGCTCCT CGCTG
• Sticky ends are produced when the enzyme makes a
  jagged cut at the recognition site
• --------------
  ACCGATTAGCGAC
  ACCGATTA GCGAC
  TGGCTAATCGCTG
  TGGC TAATCGCTC

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Restriction Mapping

• A restriction map shows the specific RE
  recognition sites

Fungal Genetics Stock Center (fgsc). Genes from
Neurospora crassa. Retrieved September 3rd, 2008
from www.fgsc.net/plasmid/plasmid.html
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Restriction Mapping

• Multiple REs used
• Each RE cuts at different locations
• Fragments move through gel according to size
• Each REs fragments creates signature bands

National Diagnostics. Retrieved Sept 3rd, 2008
from National Diagnostics.com/images
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Restriction Mapping

• Since different REs cut at different locations,
• Can deduce order by comparing fragment sets

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Restriction Mapping
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Uses of Restriction Enzymes

• Creating genome libraries.
• -Recombinant DNA technology.
• Cloning DNA molecules.
• Studying nucleotide sequence.
• Medicinal purpose
• -Insulin

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Work Cited Page.

• Campbell, N., and Reece, J. (2002). Biology
  Sixth Edition. San Francisco Benjamin Cummings.
• Nelson, L., and Cox, M. (2004). Lehringer
  Principles of Biochemistry Fourth Edition. New
  York W.H. Freeman and Company.