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Questions and Answers
What is the primary function of DNAase I?
What is the primary function of DNAase I?
- Cleave DNA at sites specified by specific DNA sequences
- Initiate synthesis from 5’ to 3’
- Digest double-stranded and single-stranded DNA
- Cleave DNA at nonspecific cleavage sites (correct)
Which enzyme is responsible for making a specific cut in DNA?
Which enzyme is responsible for making a specific cut in DNA?
- DNA Polymerase I
- Mung bean nuclease
- DNAase I
- Restriction endonucleases (correct)
What is the function of Klenow fragment?
What is the function of Klenow fragment?
- Add free nucleotides to the 3’ end of the new strand
- Initiate synthesis from 5’ to 3’ (correct)
- Cleave DNA at nonspecific cleavage sites
- Digest double-stranded and single-stranded DNA
Which enzyme possesses inherent 3’ to 5’ and 5’ to 3’ exonuclease activity?
Which enzyme possesses inherent 3’ to 5’ and 5’ to 3’ exonuclease activity?
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From which source is DNAase I isolated?
From which source is DNAase I isolated?
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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
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