Allele specific synthetic lethality between priC and dnaAts alleles at the permissive temperature of 30°C in E. coli K-12

Table 4 Strain List

Strain	priC	dnaA	dnaC	Relevant Genotype	Source or Derivation
ALO454	+	46	+	tnaA300::Tn10	Kirsten Skarstad
AQ12251	+	+	+	rnhA339::cat	T. Kogoma
CAG18442	+	+	+	thr-34::Tn10	[38]
CM740	+	5	+		Kirsten Skarstad
CM2556	+	167	+		Kirsten Skarstad
CM2733	+	602	+		Kirsten Skarstad
JC12390	+	508	+	tnaA300::Tn10	Lab stock
JC13509^a	+	+	+		[18]
JC19008	+	+	809	priA2::kan	[28]
JC19165	303	+	+		[18]
JC19257	+	+	809,820	priA2::kan	[18]
SS145	303	+	+		JC19165 -> JC13509^b
SS1091	+	+	809		JC19008 -> SS1213^c
SS1092	+	+	809,820		JC19257 -> SS1213^c
SS1099	303	+	809		JC19165 -> SS1091^b
SS1100	303	+	809,820		JC19165 -> SS1092^b
SS1201	+	508	+	tnaA300::Tn10	JC12390 -> JC13509^e
SS1213	+	+	+	thr-34::Tn10	CAG18442 -> JC13509^d
SS1504	303	508	809,820	tnaA300::Tn10	JC12390 -> SS1100^e
SS1531	303	+	+	rnhA339::cat	AQ12251 -> SS145
SS1543	303	508	+	rnhA339::cat	JC12390 -> SS1531^f
SS1738	+	+	+	tnaA300::Tn10	Lab Stock
SS1750	+	5	+	tnaA300::Tn10	SS1738 -> CM740^e
SS1751	+	167	+	tnaA300::Tn10	SS1738 -> CM2556^e
SS1752	+	602	+	tnaA300::Tn10	SS1738 -> CM2733^e
SS1793	303	167	+	tnaA300::Tn10	SS1751 -> SS145^e
SS1796	303	602	+	tnaA300::Tn10	SS1752 -> SS145^e
SS1797	303	5	+	tnaA300::Tn10	SS1750 -> SS145^e
SS1798	303	204	+	tnaA300::Tn10	WM433 -> SS145^e
SS3032	303	46	+	rnhA339::cat	ALO454 -> SS1531^f
WM433	+	204	+	tnaA300::Tn10	Mark Sutton

^aJC13509 has the following genotype: sulB103 lacMS286 Φ 80dIIlacBK1 argE3 his-4 thi-1 xyl-5 mtl-1 Sm^R T6^R. The lacMS286 Φ 80dIIlacBK1 code for two partial copies of the lac operon [39, 40].
^bSelect for kanamycin resistance on minimal media.
^cSelect for threonine prototrophy on minimal media. Screen for the presence of the dnaC mutation by the method described in [18].
^dSelect for tetracycline resistance on rich media and screen for threonine auxotrophy.
^eSelect for tetracycline resistance on rich media at 30°C and then screen for temperature sensitive growth at 42°C.
^fSelect for tetracycline resistance on rich media at 30°C and then screen for restriction site polymorphism and or backcross to test the presence of the temperature sensitive allele.

ISSN: 1471-2180