TABLE 2.
Protein mutants and primers for plasmid construction
| Plasmid IDa | Mutant | Primer nameb | Primer sequence |
|---|---|---|---|
| ppgtA3H001 | pgtα wt | ||
| ppgtA3H030 | pgtα E56A | JBp30 pgtα E56A-F | AACAAGAAAGAGGACCATGCAGCAATTCGCTTTATTAACAAGATC |
| JBp31 pgtα E56A-R | GATCTTGTTAATAAAGCGAATTGCTGCATGGTCCTCTTTCTTGTT | ||
| ppgtA3H054 | pgtα R17A, R18A | JBp123 pgt R17,18A-F | CGGGTAGTAAGGCTTGTTGACTGCGGCCTTGTTGTTAAACTGTAAGCTG |
| JBp124 pgt R17,18A-R | CAGCTTACAGTTTAACAACAAGGCCGCAGTCAACAAGCCTTACTACCCG | ||
| ppgtA3H055 | pgtα R17E, R18E | JBp125 pgt R17,18E-F | CTTCCTCGGGTAGTAAGGCTTGTTGACCTCCTCCTTGTTGTTAAACTGTAAGCTGAATGT |
| JBp126 pgt R17,18E-R | ACATTCAGCTTACAGTTTAACAACAAGGAGGAGGTCAACAAGCCTTACTACCCGAGGAAG | ||
| ppgtA3H056 | pgtα Y23A | JBp172 pgt Y23A-F | GGCCTTCCTCGGGTAGGCAGGCTTGTTGACTCGG |
| JBp173 pgt Y23A-R | CCGAGTCAACAAGCCTGCCTACCCGAGGAAGGCC | ||
| ppgtA3H057 | pgtα Y23A, R26A | JBp174 pgt Y23A, R26A-F | ACACAAGAGGGCCTTCGCCGGGTAGGCAGGCTTGTTGACTCGG |
| JBp175 pgt Y23A, R26A-R | CCGAGTCAACAAGCCTGCCTACCCGGCGAAGGCCCTCTTGTGT | ||
| ppgtA3H058 | pgtα Y23A, R26E | JBp176 pgt Y23A, R26E-F | AACACAAGAGGGCCTTCTCCGGGTAGGCAGGCTTGTTGACTCGGC |
| JBp177 pgt Y23A, R26E-R | GCCGAGTCAACAAGCCTGCCTACCCGGAGAAGGCCCTCTTGTGTT | ||
| ppgtA3H059 | pgtα R26A | JBp127 pgt R26A-F | ACACAAGAGGGCCTTCGCCGGGTAGTAAGGCTTG |
| JBp128 pgt R26A-R | CAAGCCTTACTACCCGGCGAAGGCCCTCTTGTGT | ||
| ppgtA3H060 | pgtα R26E | JBp129 pgt R26E-F | GTAACACAAGAGGGCCTTCTCCGGGTAGTAAGGCTTGTTG |
| JBp130 pgt R26E-R | CAACAAGCCTTACTACCCGGAGAAGGCCCTCTTGTGTTAC | ||
| ppgtA3H061 | pgtα R175A | JBp131 pgt R175A-F | CTTTATCCTTTCAAGCCGTGCCTTTATGGCTTGGCTGTTTTTAT |
| JBp132 pgt R175A-R | ATAAAAACAGCCAAGCCATAAAGGCACGGCTTGAAAGGATAAAG | ||
| ppgtA3H062 | pgtα R175E | JBp133 pgt R175E-F | GGGACTTTATCCTTTCAAGCCGCTCCTTTATGGCTTGGCTGTTTTTAT |
| JBp134 pgt R175E-R | ATAAAAACAGCCAAGCCATAAAGGAGCGGCTTGAAAGGATAAAGTCCC | ||
| ppgtA3H063 | pgtα R176A | JBp135 pgt R176A-F | ACTTTATCCTTTCAAGCGCTCGCTTTATGGCTTGGCTGTT |
| JBp136 pgt R176A-R | AACAGCCAAGCCATAAAGCGAGCGCTTGAAAGGATAAAGT | ||
| ppgtA3H064 | pgtα R176E | JBp137 pgt R176E-F | GACTTTATCCTTTCAAGCTCTCGCTTTATGGCTTGGCTGTTT |
| JBp138 pgt R176E-R | AAACAGCCAAGCCATAAAGCGAGAGCTTGAAAGGATAAAGTC | ||
| ppgtA3H065 | pgtα R175A, R176A | JBp139 pgt R175,176A-F | CCGGGACTTTATCCTTTCAAGCGCTGCCTTTATGGCTTGGCTGTTTTTA |
| JBp140 pgt R175,176A-R | TAAAAACAGCCAAGCCATAAAGGCAGCGCTTGAAAGGATAAAGTCCCGG | ||
| ppgtA3H066 | pgtα R175E, R176E | JBp141 pgt R175,176E-F | CACTCCGGGACTTTATCCTTTCAAGCTCCTCCTTTATGGCTTGGCTGTTTTTATCTA |
| JBp142 pgt R175,176E-R | TAGATAAAAACAGCCAAGCCATAAAGGAGGAGCTTGAAAGGATAAAGTCCCGGAGTG | ||
| phA3H032 | hu wt | JBp189 del mocr huA3H-F | GGTACCGAGAACCTGTACTTCCAATCC |
| JBp188 del mocr huA3H-R | CAGATCTACACCAGAAGAATGATGATGATG | ||
| phA3H033 | hu E56A | JBp46 hu E56A-F | GAAAACAAGAAAAAGTGCCATGCAGCAATTTGCTTTATTAACGAGATCAAGTC |
| JBp47 hu E56A-R | GACTTGATCTCGTTAATAAAGCAAATTGCTGCATGGCACTTTTTCTTGTTTTC | ||
| phA3H034 | hu R17A, R18A | JBp103 hu R17,18A-F | GGTAGTAAGGCCTTCTGAGGGCGGCCTTGTTGTTAAACTGTAAGCG |
| JBp104 hu R17,18A-R | CGCTTACAGTTTAACAACAAGGCCGCCCTCAGAAGGCCTTACTACC | ||
| phA3H038 | hu R17E, R18E | JBp105 hu R17,18E-F | CCTCGGGTAGTAAGGCCTTCTGAGCTCCTCCTTGTTGTTAAACTGTAAGCGGAA |
| JBp106 hu R17,18E-R | TTCCGCTTACAGTTTAACAACAAGGAGGAGCTCAGAAGGCCTTACTACCCGAGG | ||
| phA3H039 | hu R26A | JBp107 hu R26A-F | CACAAGAGGGCCTTCGCCGGGTAGTAAGGCCT |
| JBp108 hu R26A-R | AGGCCTTACTACCCGGCGAAGGCCCTCTTGTG | ||
| phA3H040 | hu R26E | JBp109 hu R26E-F | AACACAAGAGGGCCTTCTCCGGGTAGTAAGGCCTTC |
| JBp110 hu R26E-R | GAAGGCCTTACTACCCGGAGAAGGCCCTCTTGTGTT | ||
| phA3H041 | hu R175A | JBp111 hu R175A-F | TTTATCCTGTCAAGCCGTGCCTTTATGGCTCGACTGTTTT |
| JBp112 hu R175A-R | AAAACAGTCGAGCCATAAAGGCACGGCTTGACAGGATAAA | ||
| phA3H042 | hu R175E | JBp113 hu R175E-F | CTGCTTTATCCTGTCAAGCCGCTCCTTTATGGCTCGACTGTTTTTA |
| JBp114 hu R175E-R | TAAAAACAGTCGAGCCATAAAGGAGCGGCTTGACAGGATAAAGCAG | ||
| phA3H043 | hu R176A | JBp115 hu R176A-F | CTTTATCCTGTCAAGCGCTCGCTTTATGGCTCGACTGTTT |
| JBp116 hu R176A-R | AAACAGTCGAGCCATAAAGCGAGCGCTTGACAGGATAAAG | ||
| phA3H044 | hu R176E | JBp117 hu R176E-F | CTTTATCCTGTCAAGCTCTCGCTTTATGGCTCGACTGTTT |
| JBp118 hu R176E-R | AAACAGTCGAGCCATAAAGCGAGAGCTTGACAGGATAAAG | ||
| phA3H045 | hu R175A, R176A | JBp119 hu R175,176A-F | GGACTGCTTTATCCTGTCAAGCGCTGCCTTTATGGCTCGACTGTTTTTA |
| JBp120 hu R175,176A-R | TAAAAACAGTCGAGCCATAAAGGCAGCGCTTGACAGGATAAAGCAGTCC |
a
ID, identifier.
b
At the end of the primer name, whether the primer is a forward (F) or reverse (R) primer is indicated after the hyphen.