Table 3.
ORFs cloneda
| Organism | Gene/locus tag | UniProt accession no. | Source or reference | Primer | RE(s) used | Sequence |
|---|---|---|---|---|---|---|
| S. stipitis | XYL1 | P31867 | 24 | EY037 | XbaI | GCTCTAGAATGCCTTCTATTAAGTTGAACTCTGGTTAC |
| EY038 | ClaI | CCATCGATTTATTTCCTCTCTATAAAGCAACCTTCTTAG | ||||
| XYL2 | P22144 | 24 | EY103 | BamHI-HF | GGCGCGGATCCATGACTGCTAACCCTTCCTTGGT | |
| EY104 | XmaI | TCCCCCGGGGGATTACTCAGGGCCGTCAATGAGAC | ||||
| A. thaliana | At5g59250 | Q0WWW9 | 16 | EY052 | EcoRI | CGGAATTCATGGCTTTCGCTGTCTCGGT |
| EY004 | ClaI | CCATCGATTCACTTCAAGATTTTTGATTCAATTTCTTCC | ||||
| At5g17010 | Q6AWX0 | 16 | EY177 | BamHI-HF | GGCGCGGATCCATGGCGCTTGATCCTGAGCA | |
| EY178 | ClaI | CCGTATCCATCGATTTAGAGACATTTGGCTTCAATTTCCTCA | ||||
| C. intermedia | GXF1 | Q2MDH1 | 29 | EY175 | BamHI-HF | GGCGCGGATCCATGTCACAAGATTCGCATTCTTCT |
| EY176 | ClaI | CCGTATCCATCGATTTAAACCTGTTCGTCGGTGGCC | ||||
| GXS1 | Q2MEV7 | 29 | EY083 | BamHI-HF | CGGGATCCATGGGTTTGGAGGACAATAGAATGG | |
| EY082 | ClaI | CCATCGATTTAAACAGAAGCTTCTTCAGACATAATAGC | ||||
| D. hansenii | DEHA0D02167 | Q6BTD8 | BLAST | EY163 | XmaI | TATTCCCCCGGGATGGGTTTAGAAGATAATGCGCTTAT |
| EY164 | XhoI | CCGTGGCTCGAGTTAGACTGAAGTGGTTTCAATTTCAGT | ||||
| DEHA2B14278 | Q6BW54 | BLAST | EY087 | BamHI-HF | CCATCGATCTACGAACTAGAGTCCTTGGTATCAAC | |
| EY086 | ClaI | CGGGATCCGAGAGAAGGAGAAAAATAATACGAAATGTGG | ||||
| DEHA2A14300 | B5RSN0 | BLAST | EY181 | EcoRI | GAGTGGCGGAATTCATGTTCAATAAAATCAGATTTGGTTTCTGCA | |
| EY182 | ClaI | CCGTATCCATCGATCTACTTACTTATGCTATTTGATTCCACTTGTTC | ||||
| DEHA2F19140 | B5RUJ3 | BLAST | EY084 | BamHI-HF | CGGGATCCTTGATGTCTTCGTTATTGACCAACA | |
| EY085 | ClaI | CCATCGATTCAATTATTCGAGAGCAAAGTACGTTC | ||||
| xylHP | Q64L87 | 28 | EY167 | XmaI | TATTCCCCCGGGATGACTACTGCTGTTGGATTAGAAGATAATTCC | |
| EY168 | XhoI | CCGTGGCTCGAGTTAATCAGAATAATGTGCTTCCGAAATATCTGTTG | ||||
| E. coli | xylE | P0AGF4 | 15 | EY051 | BamHI-HF | CGGGATCCATGAATACCCAGTATAATTCCAGTTATATATTTTCG |
| EY001 | ClaI | CCATCGATTTACAGCGTAGCAGTTTGTTGTG | ||||
| S. stipitis | XUT1 | A3LY10 | NCBI | EY073 | BamHI-HF | CGGGATCCATGCACGGTGGTGGTGACGG |
| EY072 | ClaI | CCATCGATTTATTTTTCAACGTGGTAGACATCAGCCTTGC | ||||
| XUT2 | A3GIE8 | NCBI | EY061 | BamHI-HF | CGGGATCCATGAAGTATTTTCAAATCTGGAAATCAGGC | |
| EY032 | ClaI | CCATCGATCACTCAACTTCAATATGCTCGATTATTG | ||||
| XUT3 | A3GHU5 | NCBI | EY169 | XmaI | TATTCCCCCGGGATGAGAGAAGTTGGTATTCTTGATGTTGC | |
| EY170 | XhoI | CCGTGGCTCGAGTTATTCTGACATTTCAATCGAGTTGCG | ||||
| XUT4 | A3M0B9 | NCBI; intron-free version synthesized by Blue Heron Biotechnology | ||||
| XUT5 | A3LY79 | NCBI | EY068 | EcoRI | CGGAATTCATGACGGAAAGAAGCATTGGACCTT | |
| EY069 | ClaI | CCATCGATTTACTTCTTTGTATTAACAACAAAACCTTGTCTG | ||||
| XUT6 | A3M0N4 | NCBI | EY035 | EcoRI | CGGAATTCATGTCCAGTGTTGAAAAAAGTGCTGAAA | |
| EY036 | ClaI | CCATCGATTTAGCTGATGTTTTCGACATGCTCTAT | ||||
| XUT7 | A3GHF2 | NCBI | EY207 | XmaI | TATTCCCCCGGGATGATATCATCGCTTTTGGTAGC | |
| EY174 | ClaI | CCGTATCCATCGATCTAGAGTAATGTTCTTCTTGGAGACTCG | ||||
| Y. lipolytica | YALI0B06391 | Q6CFJ6 | BLAST | EY059 | EcoRI | CGGAATTCATGATTGGAAACGCTCAAATTAACCA |
| EY027 | ClaI | CCATCGATTTACAATTGAGAGGGAGGGGCG | ||||
| YALI0B01342 | Q6CG30 | BLAST | EY058 | EcoRI | CGGAATTCATGTACAAGGTCCATAACCCCTACC | |
| EY026 | ClaI | CCATCGATTTAGACATGCTCAGTTCCAGGATACT | ||||
| YALI0F06776 | Q6C2L7 | BLAST | EY023 | BamHI-HF | CGGGATCCATGTTTTCGTTAACGGGCAAACCG | |
| EY024 | ClaI | CCATCGATTTATACCGGAGGTTGAGGGAAGTC | ||||
| YALI0C06424 | Q6CCU6 | BLAST | EY171 | XmaI | TATTCCCCCGGGATGGGACTCGCTAACATCATCAACC | |
| EY172 | ClaI | CCGTATCCATCGATCTAGACAGACTCAATGTAGACGTGCTGTC | ||||
| YALI0C08943 | Q6CCJ1 | BLAST | EY210 | XmaI | TATTCCCCCGGGATGGCCATTATTGTGGCTG | |
| EY211 | ClaI | CGGTATCCATCGATCTAATCCGAATCAAATCCAGAAT | ||||
| C. neoformans | CNBC3990 | Q55VT8 | BLAST | Introns, Synthesized by Blue Heron Biotechnology | ||
| S. cerevisiae | GAL2 | P13181 | 3, 38 | EY220 | EcoRI | TATTCCGAATTCATGGCAGTTGAGGAGAA |
| EY221 | ClaI | CGGTATCCATCGATTTATTCTAGCATGGCCTTG | ||||
| HXT4 | P32466 | 15, 36 | EY194 | XmaI | TATTCCCCCGGGATGAATTCAACTCCAGATTTAATATCTCC | |
| EY208 | ClaI | CGGTATCCATCGATTTATTTCTTGCCGAACATTTTCTT | ||||
| HXT7 | P39004 | 15, 36 | EY159 | BamHI-HF | GGCGCGGATCCATGTCACAAGACGCTGCTATTGC | |
| EY180 | ClaI | CCGTATCCATCGATTTATTTGGTGCTGAACATTCTCTTGTACAATGG | ||||
| HXT13 | P39924 | 15, 36 | EY196 | XmaI | TATTCCCCCGGGATGTCTAGTGCGCAATCCTC | |
| EY209 | ClaI | CGGTATCCATCGATTCAATCAGAATTCTTTGAGAACTTC |
a
Many genes were cloned using PCR and restriction enzyme (RE) cloning. The complete list is given here, including the restriction enzymes and primers used.