TABLE 2.

Oligonucleotides used in this study

Activity and plasmid (target or product)	Oligonucleotide or oligonucleotide paira	Sequenceb (restriction endonuclease)
Deletion mutant construction
pSL217 (yscP)	Lloyd249 and Lloyd250	5′-GCC GCA TGC GCT TGC AGG CAG CTC ATC GAC-3′ (SphI)
		5′-AGC CTC CCA CTC AGT GGT GAT TTT TAT TCA TTA GGC G-3′
	Lloyd251 and Lloyd252	5′-AAA ATC ACC ACT GAG TGG GAG GCT GAA GAA TGA G-3′
		5′-GGC TCT AGA GCT GAG GTT AGC CAG AAG GAG-3′ (XbaI)
pJJ1 (yscI)	Lloyd325 and Lloyd326	5′-GCC TCT AGA GTG AAT CCG CTG AAA AGA CAC-3′ (XbaI)
		5′-CGA CAA GGT TTC AGC TAT TTC TAT GTT CGG CAT-3′
	Lloyd327 and Lloyd328	5′-ATA GAA ATA GCT GAA ACC TTG TCG AAG GGG GGG-3′
		5′-GGC CTC GAG GCC CGT AAT TTC AGA TAA AG-3′ (XhoI)
pJJ2 (yscU)	Lloyd316 and Lloyd317	5′-GCC CTC GAG CTC AAT CCT GGA CTT GAT AGT C-3′ (XhoI)
		5′-TTC GGA ATG TTG TTG CTC TGT CTT TTC TCC TCC GCT T-3′
	Lloyd318 and Lloyd333	5′-AAG ACA GAG CAA CAA CAT TCC GAA ATG TTA TAA TAG GC-3′
		5′-GCT CAC GAG CTC GCA CAG GAG AAA TAC AAT TAC C-3′ (SacI)
Antibody production
pSL303 (GST-YscF)	Lloyd222 and Lloyd244	5′-GCC GAA TTC ATG AGT AAC TTC TCT GGA TTT AC-3′ (EcoRI)
		5′-GGC CTC GAG TAA TGG AAC CTC TAT TAA GGG-3′ (XhoI)
pSL310 (GST-YscI)	Lloyd224 and Lloyd225	5′-GCC GAA TCC ATG CCG AAC ATA GAA ATA GCT C-3′ (EcoRI)
		5′-GGC CTC GAG TCA CCC CCC CTT CGA CAA GGT-3′ (XbaI)
Complementation
pSL322 (YscP)	Lloyd344 and Lloyd252	5′-GCC GCC GTC GAC GCT TGC AGG CAG CTC ATC GAC-3′ (SalI)
		5′-GGC TCT AGA GCT GAG GTT AGC CAG AAG GAG-3′ (XbaI)
pSW71 (YscI)	Lloyd325 and Lloyd328
pJJ3 (YscU)	Lloyd330 and Lloyd333	5′-GC TGA TCT AGA TAT TAA TCG CCG CTG TAT TGG C-3′ (XbaI)
		5′-GCT CAC GAG CTC GCA CAG GAG AAA TAC AAT TAC C-3′ (SacI)
Site-directed mutagenesis
pSW30 (YscU Y287G mutant form)	Lloyd544 and Lloyd545	5′-GTA ACA TTC AAA GGT ACC GAT GCC CAA G-3′
		5′-CTT GGG CAT CGG TAC CTT TGA ATG TTA C-3′
pSW31 (YscU Y317D mutant form)	Lloyd548 and Lloyd549	5′-GCC CGT GCT CTT GAT TGG GAT GCG CTC G-3′
		5′-CGA GCG CAT CCC AAT CAA GAG CAC GGG C-3′
pSW71-P77A	Lloyd359	5′-CAF TTG ATA ATG CCA ACG ACC TG-3′
pSW71-L80A	Lloyd360	5′-AAT CCC AAC GAC GCG ATG CTA ATG C-3′
pSW71-M83A	Lloyd361	5′-CGA CCT GAT GCT AGC GCA ATG GTC AC-3′
pSW71-Q84A	Lloyd362	5′-CCT GAT GCT AAT GGC ATG GTC ACT TAT CC-3′
pSW71-W85A	Lloyd363	5′-GAT GCT AAT GCA AGC GTC ACT TAT CCG-3′
pSW71-L87A	Lloyd364	5′-ATG CAA TGG TCA GCT ATC CGT ATA AC-3′
pSW71-R89D	Lloyd365	5′-TGG TCA CTT ATC GAT ATA ACA ATC CAA G-3′
pSW71-I90A	Lloyd366	5′-GTC ACT TAT CCG TGC AAC AAT CCA AGA AG-3′
pSW71-T91A	Lloyd367	5′-CTT ATC CGT ATA GCA ATC CAA GAA GAA C-3′
pSW71-Q93A	Lloyd368	5′-CGT ATA ACA ATC GCA GAA GAA CTT ATC G-3′
pSW71-E94K	Lloyd369	5′-ATA ACA ATC CAA AAA GAA CTT ATC GCC-3′
pSW71-E95K	Lloyd370	5′-ACA ATC CAA GAA AAA CTT ATC GCC AAG-3′
pSW71-L96A	Lloyd371	5′-ATC CAA GAA GAA GCT ATC GCC AAG ACT G-3′
pSW71-I97A	Lloyd372	5′-CCA AGA AGA ACT TGC CGC CAA GAC TGC CG-3′
pSW71-A98V	Lloyd373	5′-GAA CTT ATC GTC AAG ACT GCC GG-3′
pSW71-K99E	Lloyd374	5′-GAA CTT ATC GCC GAG ACT GCC GGG-3′
pSW71-T100A	Lloyd375	5′-TAT CGC CAA GGC TGC CGG GCG-3′
pSW71-G102A	Lloyd376	5′-AAG ACT GCC GCG CGA ATG AGC-3′
pSW71-Q106A	Lloyd377	5′-GGG CGA ATG AGC GCA AAT GTT GAA ACC-3′
pSW71-N107A	Lloyd378	5′-CGA ATG AGC CAA GCT GTT GAA ACC TTG-3′
pSW71-E109K	Lloyd379	5′-GCC AAA ATG TTA AAA CCT TGT CG-3′
pSW71-T110A	Lloyd380	5′-AAT GTT GAA GCC TTG TCG GAA GG-3′
pSW71-L111A	Lloyd381	5′-AAT GTT GAA ACC GCG TCG AAG GGG GG-3′
pSW71-K113E	Lloyd382	5′-ACC TTG TCG GAG GGG GGG TG-3′

^aFor oligonucleotide pairs, sequences are listed respectively.

^bRestriction sites used for subcloning are italicized. Complementary sequences for primer pairs used in overlapping PCR are underlined. Nucleotides altered via site-directed mutagenesis are in bold.