T4 Replisome Purification

Used in our papers:¹²

Contents

1. Replisome
2. Primosome
3. Assays and Labeling

1. Replisome

• DNA Polymerase GP43
• Clamp GP45
• Clamp Loader GP44/62

2. Primosome

• Helicase GP41 (and GP41 Helicase: Benkovic Purification)
• Helicase Loader GP59
• Single Stranded Binding Protein GP32
• Primase GP61

3. Assays and Labeling

• ATPase Assay (NADH-coupled ATPase activity assay for GP44/62 and GP41)
• Labeling of gp45e212c (Biotin- and Alexa Fluor 488-maleimide labeling of GP45)

Footnotes

1. Myler et al. (2016). Single-molecule imaging reveals the mechanism of Exo1 regulation by single-stranded DNA binding proteins. Proc Natl Acad Sci USA. ↩

2. Blevins et al. (2021). Characterization of the T4 gp32-ssDNA complex by native, cross-linking, and ultraviolet photodissociation mass spectrometry. Chem Sci. ↩