[Enzymatic synthesis of high-modified DNA].

Area of Science:

• Biochemistry
• Molecular Biology
• Synthetic Chemistry

Background:

• Deoxyuridine triphosphate (dUTP) derivatives are crucial for creating modified nucleic acids.
• Developing novel dUTP analogs with specific functional groups is essential for expanding DNA applications.
• Protein-like functional groups and alkene linkers offer unique modification possibilities.

Purpose of the Study:

• To synthesize and purify six novel deoxyuridine triphosphate derivatives.
• To incorporate these derivatives into single-stranded DNA using specific polymerases.
• To evaluate the utility of these modified DNA strands as templates for further DNA synthesis and aptamer development.

Main Methods:

• Chemical synthesis and purification of six deoxyuridine triphosphate derivatives.
• Enzymatic incorporation of derivatives into single-stranded DNA using KOD XL and Deep Vent polymerases.
• Assessment of DNA modification levels and template capabilities of modified DNA strands.

Main Results:

• Successfully synthesized and purified six deoxyuridine triphosphate derivatives with protein-like functional groups and varied alkene linker lengths.
• Achieved a high degree of DNA modification by incorporating these derivatives into single-stranded DNA.
• Demonstrated that the modified DNA strands can be effectively used as templates by polymerases for synthesizing unmodified DNA.

Conclusions:

• The synthesized deoxyuridine triphosphate derivatives are effective substrates for DNA modification.
• These modified DNA strands are suitable templates for synthesizing new DNA sequences.
• The derivatives hold promise for producing modified aptamers for protein targets via SELEX.