TABLE 1.

Coupling of T. kodakaraensis DNA Primase activity with Klenow Pol

Reaction mixtures (20 ml), containing 20 mm, Tris-HCl (pH 8.0), 2.5 mm DTT, 3 mm magnesium acetate, 100 mg/ml BSA, 50 mm [α-³²P]dATP (2070 cpm/pmol), 0.4 units of Klenow, 4.54 pmol of poly(dT)₂₂₁, and 50 mm NaCl were incubated for 30 min at 37 °C. Reactions with the human primase complex (p58 + p48 subunits) also included 2.5 mm ATP, where indicated. Aliquots of each reaction were plated on DEAE cellulose paper and washed three times with 0.5 m ammonium formate + 1 mm sodium pyrophosphate, dried and counted.

Additions	Protein added	dAMP incorporated
	fmol	pmol
Wild-type complex (p41-p46)	40	684
	4	72.8
	0.4	10.4
Mutant complex (m p41-p46)	40	<1
	4	<1
WT p41	40	31
	4	6.4
p46	50	<1
WT p41 + p46 (50 fmol)	40	692
	4	74.8
	0.4	7.6
Mutant p41	40	<1
Mutant p41 + p46 (50 fmol)	40	<1
Human primase complex +ATP	70	716
Human primase complex −ATP	70	<1