Table 3.
Mutated oligonucleotides for site direct mutagenesis
| oligo | forward | reverse |
|---|---|---|
| ECOR1_RWARD | GCAGAGCTCGTTTAGTGAACCGTCAGAATT | |
| XHO1_REVERSE | CTGTTCAGGAAACAGCTATGACCGCGGCCG | |
| A230T | GCGAAATTTTACTGACATTGATGATTCCTGG | CAATGTCAGTAAAATTTCGCCAGTGATTGC |
| E341D | AAGTGTGGGATCGACCTCTCTCAGGCTTAGC | GAGAGGTCGATCCCACACTTCAAAGTTGTCTC |
| L310F | AGCCAAAGCTTTCCTTCAGGATAAGGACGTA | CCTGAAGGAAAGCTTTGGCTTGAGGGCTGA |
| V269M | GATATGTTAATGATTGGCAACTTTGGCCTC | TTGCCAATCATTAACATATCTGGGTCATTCC |
| T410A | ATAAATCCCGCAGGCACTGTTTTGCTTCAG | ACAGTGCCTGCGGGATTTATGTGACTTCTTA |
| L300F | TCTAATGACTTCCGACACATCAGCCCTCAA | GATGTGTCGGAAGTCATTAGACATGAATAAAG |
| R301P | TGACCTCCCACACATCAGCCCTCAAGCCA | GGCTGATGTGTGGGAGGTCATTAGACATGAAT |
| D244H | GTATCTTGCACTGGACATCTTTTAACCAG | GATGTCCAGTGCAAGATACTCTTTATACTTT |
| Q280K | CTGGAATCAGAAAGTAACTCAGATGGCCCTC | TGAGTTACTTTCTGATTCCAGCTGAGGCCAA |
Underlined bases correspond to the mispairings with the normal alpha-galactosidase sequence used to introduce the mutations in the pCMV6-AC-AGAL vector