Fig. 6
From: Primer synthesis by a eukaryotic-like archaeal primase is independent of its Fe-S cluster
The PriX mutants are defective in primer initiation but competent in primer elongation. All the reactions contain 500 nM primase. a Primer synthesis assay for wild-type, chimera and mutant primase, in the presence of either poly(dT) ssDNA template (lanes 2–7) or poly(dT) ssDNA template and ribo(A8) primer (lanes 9–14). The reaction products were separated by PAGE under denaturing conditions and detected by phosphorimaging of the incorporated 32P-αATP. b Primer extension assay for wild-type, chimera and mutant primase, in the presence of a poly(dT) ssDNA template and 5′-labeled 32P-ribo(A8) primer. The reaction products were separated by PAGE under denaturing conditions and detected by phosphorimaging. c Primer synthesis assay for wild-type, chimera and mutant primase, in the presence of a poly(dT) ssDNA template and ApA di-nucleotide. The reaction products were separated by PAGE under denaturing conditions and detected by phosphorimaging of the incorporated 32P-αATP. Note that short oligo-ribonucleotides have been demonstrated to have aberrantly slow mobility on denaturing polyacrylamide gels40
