TABLE 1.
Primers used for mutagenesis and PCR
| Primer | Sequencea | Position (GenBank accession no.) |
|---|---|---|
| minCAT-F | 5′-ATGGGCTGCAAAAACCTGCTCGGTCTGG | 199-226 |
| minCAT-R | 5′-TCATTTGCACTGGTCCAAGTTGCTGTCAG | 2039-2067 (M26687) |
| E1-F | 5′-CTGGCAGCTCACGGAGcAAAATTTCTCCTGTAAC | 846-879 |
| E2-R | 5′-GTTACAGGAGAAATTTTgCTCCGTGAGCTGCCAG | 846-879 (M26687) |
| E3-F | 5′-AACAACGACACAAACcTGAAATACGGTGAGG | 880-910 |
| E4-R | 5′-CCTCACCGTATTTCAgGTTTGTGTCGTTGTT | 880-910 (M26687) |
| p30F1 | 5′-TATTGCCTATGAACCCCCTCCGTG | 902-925 |
| p30R1 | 5′-CTTTTCCTCTGTCTCCCTTCTAACG | 1964-1988 (J01998) |
| AK-F | 5′-GAACTCTTTGTCGACGAGAAGCAGG | 3711-3735 |
| AK-R | 5′-GGTTGAGATgCAtGGAGTAAGTCCAGTAC | 7047-7075 (J01998) |
| Pro-F | 5′-GACTTCCGGATCCAGCAC | 3237-3254 |
| Pro-R | 5′-AAACCCAGGGATCCAGAG | 3546-3563 (J01998) |
| LP5-F | 5′-AGTCCTCCGATAGACTGAG | 6-24 |
| LP6-R | 5′-CCCTTTTTATAGAGCTGGG | 8269-8287 (J01998) |
| Ak-ca1F | 5′-CCCTCGAGCGTCCCGATTGGGATTACAC | 1581-1600 |
| Ak-ca2R | 5′-GGTCGCGATCAAGTTGGGGCCTCCTTCGCT | 2113-2134 (J01998) |
| OMo-F | 5′-CCCTCGAGCGCCCAGACTGGGATTACAC | 1558-1585 |
| OMo-R | 5′-CtTTTATTAAAGATCcTTTCTGCCTCTCTAACCAAATC | 1884-1921 (J02255) |
| OAK-F | 5′-GAAAGGATCTTTAATAAAAGAGAAACTCCAG | 1917-1947 |
| OAK-R | 5′-AAATCGCGATCAAGTTGGGGCCTCCTTCGC | 2114-2134 (J01998) |
| Ak-ca1F | 5′-CCCTCGAGCGTCCCGATTGGGATTACAC | 1581-1600 |
| AK129R | 5′-GGTGGGGCTTCtGCCCGCGTTTTGGAGACCTGCTA | 1648-1682 (J01998) |
| MO129F | 5′-CCAAAACGCGGGCAGAAGCCCCACCAATTTG | 1643-1673 |
| MONCR | 5′-TCGCGATCGAGTTGGGACCTCC | 2104-2125 (J02255) |
| A110R-F | 5′-GGGATTACACCACCCAaagAGGTAGGAACCACCTAG | 1576-1611 |
| A110R-R | 5′-CTAGGTGGTTCCTACCTcttTGGGTGGTGTAATCCC | 1576-1611 (J02255) |
| H117L-F | 5′-AGGAACCACCTAGTCCtCTATCGCCAGTTGCTCCTAGC | 1599-1636 |
| H117L-R | 5′-GCTAGGAGCAACTGGCGATAGaGGACTAGGTGGTTCCT | 1599-1636 (J02255) |
| R110A-F | 5′-GGGATTACACCACCCAggcAGGTAGGAACCACCTAG | 1591-1626 |
| R110A-R | 5′-CTAGGTGGTTCCTACCTgccTGGGTGGTGTAATCCC | 1591-1626 (J01998) |
| L117H-F | 5′-AGGAACCACCTAGTTCaCTATCGCCAGTTGCTTTTAGC | 1614-1651 |
| L117H-R | 5′-GCTAAAAGCAACTGGCGATAGtGAACTAGGTGGTTCCT | 1614-1651 (J01998) |
| K214R-F | 5′-TGGTTAGAGAGGCAGAAAgGATCTTTAATAAACGAGAAACC | 1888-1928 |
| K214R-R | 5′-GGTTTCTCGTTTATTAAAGATCcTTTCTGCCTCTCTAACCA | 1888-1928 (J02255) |
| I229V-F | 5′-GAAGAAAGAGAGGAACGTgTtAGGAGAGAAACAGAGGAA | 1932-1970 |
| I229V-R | 5′-TTCCTCTGTTTCTCTCCTaAcACGTTCCTCTCTTTCTTC | 1932-1970 (J02255) |
| D244E-F | 5′-GAACGCCGTAGGACAGAGGAgGAGCAGAAAGAGAAAGAA | 1977-2015 |
| D244E-R | 5′-TTCTTTCTCTTTCTGCTCcTCCTCTGTCCTACGGCGTTC | 1977-2015 (J02255) |
a
Lowercase letters represent introduced base substitutions. Underlined bases represent restriction sites: SalI (AK-F), NsiI (AK-R), XhoI (Ak-ca1F, OMo-F), and NruI (Ak-ca2R, OAK-R, MONCR).