A thermostable d-polymerase for mirror-image PCR.

Area of Science:

• Synthetic biology
• Origin of life studies
• Biochemistry

Background:

• Biological systems exhibit homochirality, using d-nucleotides for nucleic acids and l-amino acids for proteins.
• The lack of anabolic enzymes hinders the synthesis of mirror-image biomolecules like l-DNA and l-RNA.

Purpose of the Study:

• To develop synthetic anabolic enzymes capable of building mirror-image nucleic acids.
• To create a tool for exploring the fundamental nature of biological homochirality.

Main Methods:

• Convergent synthesis strategy was employed to chemically produce a novel polymerase.
• The artificial enzyme, d-Dpo4-3C, a mutant DNA polymerase IV, was characterized for its thermostability and activity.
• Polymerase chain reactions (PCR) were utilized to test the enzyme's replication capabilities.

Main Results:

• A thermostable, mirror-image polymerase (d-Dpo4-3C) was successfully synthesized and characterized.
• The d-polymerase efficiently replicated and amplified mirror-image (l)-DNA.
• A mirror-image gene encoding the Sso7d protein was assembled using the artificial polymerase.

Conclusions:

• The development of d-Dpo4-3C provides a significant tool for synthetic biology.
• This artificial enzyme facilitates research into biological homochirality and the potential for mirror-image life forms.