Restriction Endonuclease: Difference between revisions

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Restriction endonuclease [[Enzyme|enzymes recognise]] short, specific sequences within [[DNA|DNA]], and then cut the DNA. The recognition sequence for restriction endonucleases are usually palindromic and symmetrical, this is because the enzymes are made up of 2 identical subunits. The cut leaves a 3’-hydroxyl end, and a 5’-phosphate end on each fragment.
Restriction endonuclease [[Enzyme|enzymes recognise]] short, specific sequences within [[DNA|DNA]], and then cut the DNA. The recognition sequence for restriction endonucleases are usually palindromic and symmetrical, this is because the enzymes are made up of 2 identical subunits. The cut leaves a 3’-hydroxyl end, and a 5’-phosphate end on each fragment.  


These enzymes are found in [[Bacteria|bacteria]], where they defend the cell against [[Bacteriophage|bacteriophages by]] digesting viral DNA. The bacteria protect their own DNA from digestion by modifying it. The method of this modification varies between bacterial species and strains, but the most common method is [[Methylation|methylation]] (different from [[Eukaryotic|eukaryotic methylation]]).
These enzymes are found in [[Bacteria|bacteria]], where they defend the cell against [[Bacteriophage|bacteriophages by]] digesting viral DNA. The bacteria protect their own DNA from digestion by modifying it. The method of this modification varies between bacterial species and strains, but the most common method is [[Methylation|methylation]] (different from [[Eukaryotic|eukaryotic methylation]]).  


Restriction endonucleases are used in molecular biology to break down DNA into fragments and recombine these fragments in different ways or to insert these fragments into [[Plasmids|plasmids]] for [[Recombinant DNA Technology|recombinant DNA techniques]].
Restriction endonucleases are used in molecular biology to break down DNA into fragments and recombine these fragments in different ways or to insert these fragments into [[Plasmids|plasmids]] for [[Recombinant DNA Technology|recombinant DNA techniques]].  


E.g.1 [[EcoR1|EcoRI]], found in [[E.coli|E.coli]], recognises the sequence GAATTC, and cuts between the G and the A on both strands&nbsp;<ref>BRADLEY, JR., JOHNSON, DR., POBER BR. (2006) Lecture Notes Medical Genetics. 3rd ed. Maldon, Massachusetts: Blackwell Publishing Inc</ref>.
The number of restriction fragments generated depends on two factors:


=== References ===
*The restriction enzyme used (whether it cuts at 4,6 or 8 base pairs)
*The size of the [[Genome|genome]]&nbsp;(larger genomes produce more fragments)
 
E.g.1 [[EcoR1|EcoRI]], found in [[E.coli|E.coli]], recognises the sequence GAATTC, and cuts between the G and the A on both strands&nbsp;<ref>BRADLEY, JR., JOHNSON, DR., POBER BR. (2006) Lecture Notes Medical Genetics. 3rd ed. Maldon, Massachusetts: Blackwell Publishing Inc</ref>.
 
=== References ===


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Revision as of 19:42, 19 October 2014

Restriction endonuclease enzymes recognise short, specific sequences within DNA, and then cut the DNA. The recognition sequence for restriction endonucleases are usually palindromic and symmetrical, this is because the enzymes are made up of 2 identical subunits. The cut leaves a 3’-hydroxyl end, and a 5’-phosphate end on each fragment.

These enzymes are found in bacteria, where they defend the cell against bacteriophages by digesting viral DNA. The bacteria protect their own DNA from digestion by modifying it. The method of this modification varies between bacterial species and strains, but the most common method is methylation (different from eukaryotic methylation).

Restriction endonucleases are used in molecular biology to break down DNA into fragments and recombine these fragments in different ways or to insert these fragments into plasmids for recombinant DNA techniques.

The number of restriction fragments generated depends on two factors:

  • The restriction enzyme used (whether it cuts at 4,6 or 8 base pairs)
  • The size of the genome (larger genomes produce more fragments)

E.g.1 EcoRI, found in E.coli, recognises the sequence GAATTC, and cuts between the G and the A on both strands [1].

References

  1. BRADLEY, JR., JOHNSON, DR., POBER BR. (2006) Lecture Notes Medical Genetics. 3rd ed. Maldon, Massachusetts: Blackwell Publishing Inc