Table 1 List of strains and plasmids

Strains

V. rotiferianus DAT722

DAT722

Wild-type

[11]

DAT722-Sm

DAT722; Spontaneous Sm^R mutant.

This study

MD7

DAT722-Sm; Single recombination cross-over of pVSD2 into cassette 61, Km^R

This study

d8-60a

DAT722-Sm; Δcassettes 8-60, Sm^R, Km^R

This study

d8-60b

DAT722-Sm; Δcassettes 8-60, Sm^R, Km^R

This study

d8-60b-S

DAT722-Sm; Δcassettes 8-60, Sm^R, Km^R. Spontaneous mutant of d8-60b.

This study

d8-60c

DAT722-Sm; Δcassettes 8-60, Sm^R, Km^R

This study

d8-60c-S

DAT722-Sm; Δcassettes 8-60, Sm^R, Km^R. Spontaneous mutant of d8-60c.

This study

d16-60

DAT722-Sm; Δcassettes 16-60, Sm^R, Km^R

This study

E. coli

XL1-Blue

F' proAB lacI^qZΔM15 Tn10/recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relAi, Tc^R

Stratagene

SY327 λ pir

Δ(lac pro) argE (Am) rif nalA recA56

[38]

SM10 λ pir

thi thr leu tonA lacY supE recA::RP4-2-Tc::Mu, Tc^r Km^R

[39]

Plasmids

pLOW2

Cloning vector, Km^R

[40]

pGEM-T Easy

Cloning vector, Ap^R

Promega

pMAQ1080

pGEM-T Easy carrying a 1834-bp fragment. The fragment was created using fusion PCR and consists of, in order, a 448-bp of paralog group 1 sequence, a 964-bp fragment containing aphA1 and a 410-bp paralog group 2 sequence abutted by salI restriction sites.

This study

pCVD442

Mobilisable sacB counter-selectable suicide vector, Ap^R

[41]

RK600

pJAK16

pMAQ1081

pMAQ1082

ColE1 oriV; RP4tra⁺ RP4 oriT; Cm^R; helper plasmid in triparental matings

Low copy IPTG-inducible expression vector, Cm^R

salI fragment from pMAQ1080 cloned into the unique salI site of pCVD442.

pJAK16 containing cassette 11

[42]

[43]

This study

This study