DNA Manipulation Enzymes Quiz

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What is the primary function of DNAase I?

Cleave DNA at nonspecific cleavage sites

Which enzyme is responsible for making a specific cut in DNA?

Restriction endonucleases

What is the function of Klenow fragment?

Initiate synthesis from 5’ to 3’

Which enzyme possesses inherent 3’ to 5’ and 5’ to 3’ exonuclease activity?

DNA Polymerase I

From which source is DNAase I isolated?

Bovine pancreas

Study Notes

Enzymes used in Gene Manipulation

  • Identified and isolated from different bacteria strains, commercially available as highly purified recombinant enzymes.

Classes of Enzymes used in Gene Manipulation

  • Nucleases: cut or degrade DNA molecules
  • Polymerases: copy or make new strands of DNA
  • Ligases: join pieces of DNA fragments together
  • Modifying enzymes: modify the DNA by adding or removing chemical groups
  • Topoisomerase: remove or introduce supercoils from covalently closed-circular (CCC) DNA

Nucleases

  • Exonucleases: degrade DNA at either ends
  • Exonuclease III (Exo III): cleaves dsDNA, initiates at the 3' termini of linear double-stranded DNA with 5' overhangs or blunt ends and initiates at nicked sites in double-stranded DNA
  • Catalyzes: the removal of nucleotides from linear or nicked double-stranded DNA in the 3' to 5' direction
  • Exonuclease VII (Exo VII): cleaves ssDNA

Test your knowledge on the enzymes used in gene manipulation, including nucleases, polymerases, ligases, and modifying enzymes. Learn about their functions and origins from different bacterial strains.

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