PP271, a novel bacteriophage primase-polymerase, enables Primer-Free DNA Amplification

PP271, a PrimPol from bacteriophage vB-EfaS-271, is a bona fide DNA primase that synthesizes short primers, with ADP strongly stimulating initiation.

• Why it matters: Primer-free DNA amplification needs initiating enzymes that start synthesis efficiently while limiting primase-derived errors and maintaining broad sequence compatibility.
• What they did: PP271 was characterized as an AEP-superfamily PrimPol with a conserved catalytic core and PriCT domain, and its priming activity was tested for ADP stimulation and relaxed priming-site specificity. TruePrime amplification was then used with Thermus thermophilus PrimPol and Phi29 DNA polymerase, and PP271-generated primers were assessed for Phi29 extension.
• The result: PP271 primers are efficiently extended by Phi29 DNA polymerase, and the short, less sequence-restricted primers are expected to reduce primase-derived errors while supporting DNA synthesis across more sequence contexts.

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