List of 17 Enzymes with their function that are used in recombinant DNA technology /Genetic Engineering
List of 17 Enzymes with their function that are used in recombinant DNA technology /Genetic Engineering
1. Alkaline phosphatase: This enzyme helps to remove the phosphate
groups from 5’-ends of double and single standard DNA and RNA. It also helps prevent
re-circularization during cloning and dephosphorylate DNA before Kinase
labeling protocols.
2. Terminal Transferase: Enzyme very useful in homopolymer tailing.
It helps to add nucleotides to the 3’ends of DNA or RNA. This enzyme is used as
a template for a primer in subsequent PCR.
3. Bal 31 nuclease: In DNA technology this enzyme is used to
degrade both 3’ and 5’ termini of duplex DNA without generating any internal
scissions. In short, this enzyme is used for the progressive shortening of DNA.
4. S1nuclease:
It is an endonuclease enzyme that helps to degrade single-standard DNA and RNA
but can’t degrade double-standard DNA or RNA-DNA hybrids.
5. DNA Ligase:
In DNA replication, the DNA ligase enzyme helps to join DNA molecules by
forming phosphodiester linkages between DNA segments that help to synthesize
DNA to form a seamless strand. This enzyme is also used in DNA cloning for the
same process.
6. Taq DNA Polymerase: Taq DNA polymerase enzyme used in PCR i.e.
Polymerase Chain Reaction to recruit new nucleotides to single strands of DNA
in the denaturation step that is taking place in a PCR reaction that helps to
complete DNA replication to make DNA copies.
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7. DNA polymerase 1: This enzyme is coded by the pol1 gene that
helps in prokaryotic DNA replication. DNA polymerase helps to synthesize the
DNA complementary to a DNA template by removing RNA primer during the
processing of DNA after replication. It also fills the gap.
8. RNase H:
RNase H is known as the Ribonuclease H enzyme. This is an endoribonuclease
enzyme that helps to cleave and digest the RNA strand of RNA-DNA heteroduplex
(RNA-DNA hybrids). The purpose of the RNase H enzyme is to form double-strand
cDNA.
9. DNase 1:
It is a type of recombinant endonuclease enzyme that helps to produce
single-standard nicks in DNA.
10. RNase A:
RNase A is a Ribonuclease enzyme that helps in cleavage and digests RNA (and
not DNA). This enzyme unwinds the RNA helix by complexing with single-standard
RNA. The major role of this enzyme is in RNA metabolism and regulation of gene
expression.
11. Exonuclease III: This enzyme has exonucleolytic activity. It
helps to remove nucleotides from the 3’-end of DNA.
12. Reverse transcriptase: It helps to synthesize DNA from RNA to
produce complementary DNA (cDNA) libraries from variously expressed mRNAs. This
enzyme is also used in RT PCR.
13. Polynucleotide Kinase: PNK (Polynucleotide kinase) that helps to
transfer a phosphate group from ATP (Adenosine triphosphate) to 5’OH ends of
DNA or RNA. Sometimes they show phosphate activities.
14. Restriction Enzymes: The restriction enzyme cuts double-standard
DNA with a specific recognition site. It is a type of protein that is isolated
from bacteria.
15. Type I Restriction endonuclease: This enzyme requires ATP to cut DNA about
1000bp away from its restriction site. It shows both methylation and
endonuclease activity.
16. Type II Restriction endonuclease: This enzyme doesn't require ATP to cut DNA. It
cuts DNA at the restriction site.
17. Type III Restriction endonuclease: This enzyme requires ATP to cut DNA about 25bp
away from the restriction site.
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