. 2004 Nov;168(3):1145–1157. doi: 10.1534/genetics.104.032136

TABLE 3.

IS981-mediated mutations detected in single-clone representatives of the evolvedL. lactis populations

		First detected using
Population	Band change^a	PCR^b	Southern hybridization^c	Position^d (kb)	Mutational event
Growth
G/S1	Loss 1.8 kb	ND	800	1216	Deletion of >11.3 kb at left flank of IS981 E, affecting at least ymbCDG and citRCDEF (Figure 2)
G/S3	Gain 2.5 kb	300	800	1969	Insertion of IS981 107 bp upstream of ytgF (Figure 3A)
Shift 6.6 kb	500^e	400	1277	Deletion of 5.1 kb at right flank of IS981 G, affecting ymhC, amyL, lctO, and aroH (Figure 3B)
G/NS1	Shift 8.7 kb	300^f	Never	92	Deletion of 1002 bp between IS981A and IS981B, affecting yajF (Figure 4)
Starvation
NG/S1	Shift 8.7 kb	>14 days	ND	92	Deletion of ∼800 bp between IS981 A and IS981 B, affecting yajF (Figure 4)
NG/S2	Gain 3.2 kb	>14 days	ND	896	Insertion of IS981 into mleR (Figure 5)
NG/NS1	Loss 6.6 kb	ND	ND	1277	Deletion of >10 kb at right flank of IS981 G (Figure 3B)

G, growth; NG, starvation; S, shaken; NS, nonshaken; ND, not determined.

^{^a}

Detected on genomic HindIII digestions hybridized with a probe against IS981 (see materials and methods).

^{^b}

Time in generations (growing populations) or days (starving populations), based on PCR analysis of population samples isolated at 100-generation intervals; for starving populations, only a single sample taken after 14 days of evolution was available for the timing of the mutational events.

^{^c}

Time in generations, using Southern hybridizations of population samples isolated at 200-generation intervals.

^{^d}

Chromosomal coordinates of (one of the) IS981 element(s) involved in the mutation.

^{^e}

The 5.1-kb deletion in G/S3 probably happened via at least one intermediate step, as a (probably smaller) deletion was visible on Southern hybridizations from generation 400, while PCR amplification of the final 5.1-kb deletion end point was possible only with samples from generation 500 and later.

^{^f}

The deletion between existing elements IS981A and IS981B happened via at least one intermediate step (a deletion of 878 bp) in this population, which was detectable from generation 300 to 1000 (see Figure 4B).