Table 1. Mutagenic Primers Utilized To Generate EIZS Mutantsa.
| mutation | F or Rb | 5′–3′ sequence |
|---|---|---|
| L72A | F | GTC TGT GCT ACA CGG ACg ctA TGG CGG GCT ACT ACC TG |
| R | CAG GTA GTA GCC CGC CAT agc GTC CGT GTA GCA CAG AC | |
| L72V | F | CTG TGC TAC ACG GAC gtg ATG GCG GGC TAC TAC |
| R | GTA GTA GCC CGC CAT cac GTC CGT GTA GCA CAG | |
| L72M | F | GTC TGT GCT ACA CGG ACa tgA TGG CGG GCT ACT ACC TG |
| R | CAG GTA GTA GCC CGC CAT cat GTC CGT GTA GCA CAG AC | |
| F95A | F | GAC TAC AGC GCG TGG gca TTC GTC TGG GAC GAC |
| R | GTC GTC CCA GAC GAA tgc CCA CGC GCT GTA GTC | |
| F95V | F | GAT AGC GGA CTA CAG CGC GTG Ggt tTT TGT CTG GGA CGA CCG TCA C |
| R | GTG ACG GTC GTC CCA GAC AAA aac CCA CGC GCT GTA GTC CGC TAT C | |
| F95M | F | GAT AGC GGA CTA CAG CGC GTG Gat gTT TGT CTG GGA CGA CCG TCA C |
| R | GTG ACG GTC GTC CCA GAC AAA cat CCA CGC GCT GTA GTC CGC TAT C | |
| F95H | F | GAT AGC GGA CTA CAG CGC GTG Gca tTT TGT CTG GGA CGA CCG TCA C |
| R | GTG ACG GTC GTC CCA GAC AAA atg CCA CGC GCT GTA GTC CGC TAT C | |
| F96V | F | GAT AGC GGA CTA CAG CGC GTG gtt CGT TGT CTG GGA CGA CCG TCA C |
| R | GTG ACG GTC GTC CCA GAC AAC GAA cca CGC GCT GTA GTC CGC TAT C | |
| F96L | F | GAC TAC AGC GCG TGG TTC tta GTC TGG GAC GAC CGT CAC |
| R | GTG ACG GTC GTC CCA GAC taa GAA CCA CGC GCT GTA GTC | |
| F96Y | F | GAT AGC GGA CTA CAG CGC GTG GTT Cta tGT CTG GGA CGA CCG TCA C |
| R | GTG ACG GTC GTC CCA GAC ata GAA CCA CGC GCT GTA GTC CGC TAT C | |
| F96W | F | GAC TAC AGC GCG TGG TTC tgg GTC TGG GAC GAC CGT CAC |
| R | GTG ACG GTC GTC CCA GAC cca GAA CCA CGC GCT GTA GTC | |
| F198V | F | GAA CTG CGC CGG CTC ACG gtt GCT CAC TGG ATC TGG AC |
| R | GTC CAG ATC CAG TGA GCa acC GTG AGC CGG CGC AGT TC | |
| F198L | F | CTC GAA CTG CGC CGG CTC ACG tta GCG CAC TGG ATC TGG ACC GAC |
| R | GTC GGT CCA GAT CCA GTG CGC taa CGT GAG CCG GCG CAG TTC GAG | |
| F198Y | F | GTA CCT CGA ACT GCG CCG GCT CAC Gta tGC ACA CTG GAT CTG GAC CGA C |
| R | GTC GGT CCA GAT CCA GTG TGC ata CGT GAG CCG GCG CAG TTC GAG GTA C | |
| A236G | F | GTC AGG AAT TCg gcG CCT GGT ACA AC |
| R | GTT GTA CCA GGC gcc GAA TTC CTG AC | |
| A236F | F | GCG TTG CTG AGT CAG GAA TTC ttt GCC TGG TAC AAC GAC CTC TGC |
| R | GCA GAG GTC GTT GTA CCA GGC aaa GAA TTC CTG ACT CAG CAA CGC | |
| A236M | F | GCG TTG CTG AGT CAG GAA TTC atg GCC TGG TAC AAC GAC CTC TGC |
| R | GCA GAG GTC GTT GTA CCA GGC cat GAA TTC CTG ACT CAG CAA CGC | |
| W325F | F | GGC AAT ATG CGG AAC TTC ttc AGT TCC GTC TAC TG |
| R | CAG TAG ACG GAA CTG AAg aaG TTC CGC ATA TTG CC | |
| V329A | F | CTG GTT CAG TTC Cgc gTA CTG GTT CCA CC |
| R | GGT GGA ACC AGT Acg cGG AAC TGA ACC AG | |
| V329G | F | CTG GTT CAG TTC Cgg tTA CTG GTT CCA CCA CGA GTC |
| R | GAC TCG TGG TGG AAC CAG TAa ccG GAA CTG AAC CAG | |
| V329I | F | CTG GTT CAG TTC Cat tTA CTG GTT CCA CCA CGA GTC |
| R | GAC TCG TGG TGG AAC CAG TAa atG GAA CTG AAC CAG | |
| V329M | F | CTG GTT CAG TTC Cat gTA CTG GTT CCA CCA CGA GTC |
| R | GAC TCG TGG TGG AAC CAG TAc atG GAA CTG AAC CAG | |
| F332A | F | CAG TTC CGT CTA CTG Ggc aCA CCA CGA GTC CGG C |
| R | GCC GGA CTC GTG GTG tgc CCA GTA GAC GGA ACT G |
a
Lowercase letters represent the introduced mutant codon.
b
F, forward primer; R, reverse primer.