Table 2.
Primers used for construction of truncated bifunctional pfDHFR-TS
| Name | Length (bases) | Sequence (5′ ⟶ 3′) | Utilities and description |
|---|---|---|---|
| Δ229-277 Sense |
60 |
ACAACATTGGATTTTATCATTTATAAGAAATATGAAAATGATGATGATGATGAAGAAGAA |
Sense strand PCR primer for the construction of pfDHFR-TSΔ229-277. Lys228 (italic) was followed by Tyr278(bold) |
| Δ229-277 Antisense |
60 |
TTCTTCTTCATCATCATCATCATTTTCATATTTCTTATAAATGATAAAATCCAATGTTGT |
Antisense strand PCR primer for the construction of pfDHFR-TSΔ229-277 |
| Δ229-276 Sense |
60 |
ACAACATTGGATTTTATCATTTATAAGAAAAATTATGAAAATGATGATGATGATGAAGAA |
Sense strand PCR primer for the construction of pfDHFR-TSΔ229-276. Lys228 (italic) was followed by Asn277(bold) |
| Δ229-276 Antisense |
60 |
TTCTTCATCATCATCATCATTTTCATAATTTTTCTTATAAATGATAAAATCCAATGTTGT |
Antisense strand PCR primer to amplify pfDHFR-TSΔ229-276 |
| Δ229-275 Sense |
60 |
ACAACATTGGATTTTATCATTTATAAGAAAATTAATTATGAAAATGATGATGATGATGAA |
Sense strand PCR primer for the construction of pfDHFR-TSΔ229-275. Lys228 (italic) was followed by Ile276(bold) |
| Δ229-275 Antisense |
60 |
TTCATCATCATCATCATTTTCATAATTAATTTTCTTATAAATGATAAAATCCAATGTTGT |
Antisense strand PCR primer to amplify pfDHFR-TSΔ229-275 |
| Δ229-274 Sense |
60 |
ACAACATTGGATTTTATCATTTATAAGAAAAAAATTAATTATGAAAATGATGATGATGAT |
Sense strand PCR primer for the construction of pfDHFR-TSΔ229-274. Lys228 (italic) was followed by Lys275(bold) |
| Δ229-274 Antisense |
60 |
ATCATCATCATCATTTTCATAATTAATTTTTTTCTTATAAATGATAAAATCCAATGTTGT |
Antisense strand PCR primer to amplify pfDHFR-TSΔ229-274 |
| Δ229-271 Sense |
60 |
ACAACATTGGATTTTATCATTTATAAGAAAGACAAATATAAAATTAATTATGAAAATGAT |
Sense strand PCR primer for the construction of pfDHFR-TSΔ229-271. Lys228 (italic) was followed by Asp272(bold) |
| Δ229-271 Antisense |
60 |
ATCATTTTCATAATTAATTTTATATTTGTCTTTCTTATAAATGATAAAATCCAATGTTGT |
Antisense strand PCR primer to amplify pfDHFR-TSΔ229-271 |
| Δ229-265 Sense |
60 |
ACAACATTGGATTTTATCATTTATAAGAAAGAATTTTACAAAAATGTAGACAAATATAAA |
Sense strand PCR primer for the construction of pfDHFR-TSΔ229-265. Lys228 (italic) was followed by Glu266(bold) |
| Δ229-265 Antisense | 60 | TTTATATTTGTCTACATTTTTGTAAAATTCTTTCTTATAAATGATAAAATCCAATGTTGT | Antisense strand PCR primer to amplify pfDHFR-TSΔ229-265 |