TABLE 1.
Primers used in this studya
| Primer name | Position no. | Sequence | Function |
|---|---|---|---|
| lef4Cm-F | 76727-76775 | GTACGACGAAAACGGCTTTCGCACTCGTATACCTATTCAGAGCGCTTGCTGTGTAGGCTGGAGCTGCTTCG | lef-4 KO |
| lef4Cm-R | 77981-77933 | CACGATTCGGTCGCGACGATGTTTCAACACGTTTATTGTCGTGTCCGTGCATATGAATATCCTCCTTA | lef-4 KO |
| ptpZEO-F | 509-558 | CCCGCGCGTTGGCAAACTATTTACAATGCGGCCAAGTTATAAAAGATTCTGTTGACAATTAATCATCGGC | ptp/bvp KO |
| ptpZEO-R | 1080-1031 | GGTTCGCTGGAAGAAGCGCAACAATTTGAAAATAGGATAAAGTATTTGTTCCCGGGAATTCTCAGTCCTGC | ptp/bvp KO |
| A | 373-394 | GTGTGCAAAACATGACATCAGC | Confirm ptp/bvp KO |
| B | 1265-1285 | ACATCCGGTGTTGTGCGTGAC | Confirm ptp/bvp KO |
| C | 76504-76525 | GACACGATGGACGCGAAAATGC | Confirm lef-4 KO |
| D | 78286-78306 | CGCCGCCTTCTCCTATTCCTC | Confirm lef-4 KO |
| ptpR-F | 287-303 | gctactagtGCAAAACATGACATCAG | bvp/ptp repair |
| ptpR-R | 988-1006 | TTATTAAATTAATAAATCTTGAAC | bvp/ptp repair |
| p6.9polyA-F | 86478-86494 | gctactagtGGAAAAACATACCAGCA | Add polyA signal to bvp/ptp |
| p6.9polyA-R | 86695-86711 | atccccgggCAACCAGACATTCCACAC | Add polyA signal to bvp/ptp |
| lef4R-F | 76238-76254 | atcgtcgacCGGCTTGCATCATACTG | lef-4 repair |
| lef4R-R | 77970-77987 | atccccgggATTTGGCACGATTCGGTC | lef-4 repair |
| vp39polyA- F | 75276-75295 | CGCTGCATTTAATCACATCC | Add polyA signal to lef-4 |
| vp39polyA-R | 75513-75534 | gctactagtAAATGGAGTTTGTTAAATTG | Add polyA signal to lef-4 |
| D1/D12 E192/194A-F | GGCCTAATACATCTCTGGcAATAGcATTCACACCTAGAGAC | Construct RTPase− D1/D12 |
a
Nucleotides added for cloning purposes are shown in lowercase type, restriction enzyme sites are underlined, mutagenic residues are indicated by lowercase bold type. Primers for lef-4 mutagenesis have been described previously (28). KO, knockout.