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OBJECTIVES
What is DNA
Ligase?
Steps/techniqu
e of DNA Ligase
Application Importance
Advantages Disadvantag
es
VIDEO CLIP
WHAT IS DNA LIGASE?
DNA ligase is a type of
enzyme that facilitates the
joining of DNA strands
together by catalyzing the
formation of a
phosphodiester bond.
DNA ligase is also called
“molecular glue”. During
DNA replication, It attaches
the Okazaki fragments
formed from the lagging
strand during the
replication of DNA.
DNA ligase works by catalyzing the
formation of a phosphodiester bond
between nucleotides on one strand
of a double stranded DNA molecule.​
HOW DOES DNA LIGASE
WORK?
DNA ligases play an essential role
in maintaining genomic integrity by
joining breaks in the phosphodiester
backbone of DNA that occur during
replication and recombination, and as a
consequence of DNA damage and its repair.
IMPORTANCE
STEPS OF DNA LIGASE
• In the first step, the DNA fragments or DNA segments or Okazaki
fragments are reorganized with their active sites.
• The lysine residue of the active site of the Ligase gets adenylated
with the addition of AMP. The release of pyrophosphate
accompanies adenylation of Ligase.
• Ligase transfer the AMP to the 5′ phosphate end of the donor
with the formation of the pyrophosphate bond.
• The 5′ phosphate end of the donor is linked to the 3′ hydroxyl
end of the receptor with phosphodiester bond formation.
APPLICATION
• DNA ligases play a key role in DNA cloning and in NGS library
preparation.
• In cloning applications, DNA ligases are largely used to bond the ends of a
DNA fragment to the ends of a plasmid or vector. This vector will
ultimately be transformed into cells of interest where it will recombine
with the cell’s wild-type DNA, generating a mutant.
APPLICATION
• DNA ligases play a key role in DNA cloning and in NGS library
preparation.
• In cloning applications, DNA ligases are largely used to bond the ends of a
DNA fragment to the ends of a plasmid or vector. This vector will
ultimately be transformed into cells of interest where it will recombine
with the cell’s wild-type DNA, generating a mutant.
ADVANTAGES
• DNA REPAIR: DNA Ligase plays a crucial role in
repairing DNA damage; such as nicks and gaps in the
DNA molecule.
• Recombinant DNA Technology: DNA Ligase is widely
used in molecular biology techniques, such as cloning and
recombinant DNA technology.
• Without DNA Ligase, the process of mutation will not
complete
ADVANTAGES
• DNA REPAIR: DNA Ligase plays a
crucial role in repairing DNA
damage; such as nicks and gaps in
the DNA molecule.
• Recombinant DNA Technology: DNA
Ligase is widely used in molecular
biology techniques, such as cloning
and recombinant DNA technology.
• Without DNA Ligase, the process of
mutation will not be complete.
DISADVANTAGES
• Although DNA Ligase is highly accurate in
sealing DNA strands, there is still a
possibility of errors such as misligation,
resulting in mutations or rearrangements in
the DNA sequence.
• Size Limitations: DNA Ligases may have
limitations on the size of DNA fragments
they can effectively ligate.
• Specificity Issues: While DNA Ligases
generally have high specificity for joining
DNA strands, they may sometimes ligate
DNA fragments with incompatible ends or
ligate non-specific DNA molecules, leading to
undesirable outcomes in molecular biology
experiments.
THANK YOU
FOR LISTENING!

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DNA LIGASE BIOTECHNOLOGY BIOLOGY STUDY OF LIFE

  • 1.
  • 2. OBJECTIVES What is DNA Ligase? Steps/techniqu e of DNA Ligase Application Importance Advantages Disadvantag es
  • 4. WHAT IS DNA LIGASE? DNA ligase is a type of enzyme that facilitates the joining of DNA strands together by catalyzing the formation of a phosphodiester bond. DNA ligase is also called “molecular glue”. During DNA replication, It attaches the Okazaki fragments formed from the lagging strand during the replication of DNA.
  • 5. DNA ligase works by catalyzing the formation of a phosphodiester bond between nucleotides on one strand of a double stranded DNA molecule.​ HOW DOES DNA LIGASE WORK?
  • 6. DNA ligases play an essential role in maintaining genomic integrity by joining breaks in the phosphodiester backbone of DNA that occur during replication and recombination, and as a consequence of DNA damage and its repair. IMPORTANCE
  • 7. STEPS OF DNA LIGASE • In the first step, the DNA fragments or DNA segments or Okazaki fragments are reorganized with their active sites. • The lysine residue of the active site of the Ligase gets adenylated with the addition of AMP. The release of pyrophosphate accompanies adenylation of Ligase. • Ligase transfer the AMP to the 5′ phosphate end of the donor with the formation of the pyrophosphate bond. • The 5′ phosphate end of the donor is linked to the 3′ hydroxyl end of the receptor with phosphodiester bond formation.
  • 8. APPLICATION • DNA ligases play a key role in DNA cloning and in NGS library preparation. • In cloning applications, DNA ligases are largely used to bond the ends of a DNA fragment to the ends of a plasmid or vector. This vector will ultimately be transformed into cells of interest where it will recombine with the cell’s wild-type DNA, generating a mutant.
  • 9. APPLICATION • DNA ligases play a key role in DNA cloning and in NGS library preparation. • In cloning applications, DNA ligases are largely used to bond the ends of a DNA fragment to the ends of a plasmid or vector. This vector will ultimately be transformed into cells of interest where it will recombine with the cell’s wild-type DNA, generating a mutant.
  • 10. ADVANTAGES • DNA REPAIR: DNA Ligase plays a crucial role in repairing DNA damage; such as nicks and gaps in the DNA molecule. • Recombinant DNA Technology: DNA Ligase is widely used in molecular biology techniques, such as cloning and recombinant DNA technology. • Without DNA Ligase, the process of mutation will not complete
  • 11. ADVANTAGES • DNA REPAIR: DNA Ligase plays a crucial role in repairing DNA damage; such as nicks and gaps in the DNA molecule. • Recombinant DNA Technology: DNA Ligase is widely used in molecular biology techniques, such as cloning and recombinant DNA technology. • Without DNA Ligase, the process of mutation will not be complete. DISADVANTAGES • Although DNA Ligase is highly accurate in sealing DNA strands, there is still a possibility of errors such as misligation, resulting in mutations or rearrangements in the DNA sequence. • Size Limitations: DNA Ligases may have limitations on the size of DNA fragments they can effectively ligate. • Specificity Issues: While DNA Ligases generally have high specificity for joining DNA strands, they may sometimes ligate DNA fragments with incompatible ends or ligate non-specific DNA molecules, leading to undesirable outcomes in molecular biology experiments.