Table 2.
Oligonucleotides used in this study Uppercase letters indicate identity to wild-type sequence. Oligonucleotides 784 and 786 contain a codon 252 mutation (GATAsp to GCTAla) and a silent mutation at codon 248 (TCASer to TCTSer) that creates a diagnostic DraI site. Oligonucleotides 783, 785, 798, and 799 contain a mutation at codon 255 (GAAGlu to GCGAla) that creates MluI and AflIII sites. Oligonucleotide 781 contains a BamHI site. Oligonucleotide 782 contains a SpeI site. Oligonucleotides 479 and 480 contain a BamHI site.
| Oligonucleotide | Name | Sequence |
|---|---|---|
| 168 | TUB1-182U(+) | CACCGTAGTTTCCGGCCG |
| 378 | TUB3-172U | GAGCAGAGCATATCGTCC |
| 479 | KStub3-01 | ccgccatggagATGAGAGAGGTCATTAGT |
| 480 | KStub3-02 | gcgccatggTTAGAACTCCTCAGCGTA |
| 781 | KAtub1-01 | cggggatccATGAGAGAAGTTATTAGTA |
| 782 | KAtub1-02 | ggactagtAGAAAGGATAAGGAGGTTG |
| 783 | KAtub1-E255A | AATGTAGATTTGAACGcgTTTCAAACCAATTTGG |
| 785 | KAtub1-E255Arev | CCAAATTGGTTTGAAAcgCGTTCAAATCTACATT |
| 784 | KAtub1-D252A | GATTCGACGGTTCtTTAAATGTAGcTTTGAACGAATTTCAAAC |
| 786 | KAtub1-D252rev | GTTTGAAATTCGTTCAAAgCTACATTTAAaGAACCGTCGAATC |
| 798 | KAtub3E255A-F | AACGcgTTTCAGACCAACTTGGTA |
| 799 | KAtub3E255A-R | GTCTGAAAcgCGTTCAAATCCAC |
| 1068 | KAtub1-03 | AATATCACCGTAGTTTCCGGC |
| 1069 | KAtub1-04 | AGAAAGGATAAGGAGGTTGGG |