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Research Article

Sulfolobus Mutants, Generated via PCR Products, Which Lack Putative Enzymes of UV Photoproduct Repair

Table 1

Strains, plasmids, and primers.
DesignationGenotypeSource or reference
S. acidocaldarius strains
DG185Wild-typeObtained as ATCC 33909
MR31 𝑝 𝑦 𝑟 𝐸 1 3 1 S a c (18-bp internal deletion)[21]
LR10Saci_1227:: 𝑝 𝑦 𝑟 𝐸 + S s o Pyr+ transformant of MR31 (see text)
LR12Saci_1096:: 𝑝 𝑦 𝑟 𝐸 + S s o Pyr+ transformant of MR31 (see text)
DG251Saci_1427:: 𝑝 𝑦 𝑟 𝐸 + S s o Pyr+ transformant of MR31 (see text)
DG260Saci_1227:: 𝑝 𝑦 𝑟 𝐸 − S s o ,Spontaneous FOA-resistant LR10
𝑝 𝑦 𝑟 𝐸 1 3 1 S a c
DG262Saci_1096:: 𝑝 𝑦 𝑟 𝐸 − S s o ,Spontaneous FOA-resistant LR12
𝑝 𝑦 𝑟 𝐸 1 3 1 S a c
CS1Saci_1227:: 𝑝 𝑦 𝑟 𝐸 − S s o DG260 transformed to 𝑝 𝑦 𝑟 𝐸 + S a c
CS3Saci_1096:: 𝑝 𝑦 𝑟 𝐸 − S s o DG262 transformed to 𝑝 𝑦 𝑟 𝐸 + S a c
CS1mESaci_1227:: 𝑝 𝑦 𝑟 𝐸 − S s o Spontaneous FOA-resistant CS1
𝑝 𝑦 𝑟 𝐸 S a c   Δ G199
CS1m15Saci_1227:: 𝑝 𝑦 𝑟 𝐸 − S s o    Spontaneous FOA-resistant CS1
𝑝 𝑦 𝑟 𝐸 S a c   A335T
JDS8 𝑝 𝑦 𝑟 𝐸 S a c   Δ G199Spontaneous FOA-resistant DG185 (lab collection)
JDS28 𝑝 𝑦 𝑟 𝐸 S a c   A335TSpontaneous FOA-resistant DG185 (lab collection)
CS3m65Saci_1096:: 𝑝 𝑦 𝑟 𝐸 − S s o  Spontaneous FOA-resistant CS3
  𝑝 𝑦 𝑟 𝐸 S a c    Δ C78
CS3m78Saci_1096:: 𝑝 𝑦 𝑟 𝐸 − S s o   Spontaneous FOA-resistant CS3
𝑝 𝑦 𝑟 𝐸 S a c   G203A
DGm18 𝑝 𝑦 𝑟 𝐸 S a c    Δ C78Spontaneous FOA-resistant DG185 (lab collection)
DGm23 𝑝 𝑦 𝑟 𝐸 S a c   G203ASpontaneous FOA-resistant DG185 (lab collection)
E. coli plasmids
pMJ03SSV1-derived Sulfolobus shuttle vector[24]
pPCBE123′ portion of Saci_1427 in MCS of pUC19Phil Clark (unpublished)
pLK3a 𝑝 𝑦 𝑟 𝐸 S s o in MCS (XbaI site) of pNEB193This work
pLK4a 𝑝 𝑦 𝑟 𝐸 S s o in trpC insert of pPCBE12This work
PCR primers
SsoPEAvrKpnr1GCA GCC TAG GTA CCG ATA TGA GAG AGG TTT ATC CAT TGC
SsoPEFKpnAvrf1GCA CCT AGG ACG TGT CTT AAT CTC ACA AAG CCC TTA T
SacPhr::cassf1
CAACGAGACGAGAAAATGAAGGAAAATGCCTTGAATAAAGGAATTAAATTTACCGCACAGAGCGTAAGGAGAA
SacPhr::cassr1
ATCCACTAATTTTGTCGCAAAATACCTCTCTCCCAGTCTCCAGTCGACAATTATAGTCCTGTCGGGTTTCGCCA
Saci1096 DTf3
CACTTAGAATTATTCACTGTTGAGAGTAGGTTACGTATCCAAGTCTAGGGGTACCACGTGTCTTAATCTC
Saci1096 DTr2
GATTCACCTTAATAAATCCTCTAATCCAGTTTGTTTAACTCCTAATGCAGCTGGCACGACAGGTTTC
Saci1227fAAGAGAGACCACCGGTCTGAATGT
Saci1227rCCT TGGTCATGCCTTCGTGCTAAA
Saci_1227internalFCACCATTTTACACTAAAGCAAGAG
Saci_1227internalRCGACCAACATTCTCACTCTTC
NXSaci1096fGAACGCCGGCTCGAGCAAGAGGGTCAAGTCGATAATTGG
NXSaci1096rGAGGCCGGCTCGAGGGGCGTTTGGTATACTGTTCTATC
Saci_1096internalFGAGTGCTTAAAGTCTCTTCCTC
Saci_1096internalRCTTATCCTTCACCTCAAGCATG