Primers used for mutagenesis
| Primer . | Nucleotide sequence, 5′ to 3′ . | Position in Nramp2 . |
|---|---|---|
| pCB6 F | aatgggcggtaggcgtgta | 848-866 of pCB6 |
| pCB6 R | aggccaggagaggcactgg | 1006-1034 of pCB6 |
| N2 Asp86Ala F | cctacctagCcccaggaaac | 248-267 |
| N2 Arg119Ala F | ctgctgctccagGCccttgcag | 346-367 |
| N2 Arg146Ala F | caaggtcccaGCgatcatcctgtg | 426-449 |
| N2 Glu154Ala F | ctgatggtggCgttggcaatca | 451-472 |
| N2 Asp161Ala F | cattggttctgCcatgcaggaagtc | 471-495 |
| N2 Gly184Asp F | ccctgtgggAcggagtcctca | 542-563 |
| N2 Gly185Arg F | gggtccccctgtggggcaGagtcctcatcaccatc | 536-570 |
| N2 Asp192Ala F | caccatcgcagCcacttttgtg | 564-585 |
| N2 Glu225Ala F | gtttggatatgCgtacattac | 663-684 |
| N2 His267Ala F | atcatgccgGCcaacatgtacct | 790-825 |
| N2 His267Cys F | atcatgccgTGcaacatgtacct | 790-825 |
| N2 His267Arg F | atcatgccgcGGaacatgtacct | 790-825 |
| N2 His272Ala F | aacatgtacctgGCttctgcctta | 802-825 |
| N2 His272Cys F | aacatgtacctgTGttctgcctta | 802-825 |
| N2 His272Arg F | atgtacctgcGttctgcctta | 805-825 |
| N2 His267/272Ala F | atcatgccgGCcaacatgtacctgGCttctgcgtta | 790-825 |
| N2 His267/272Cys F | atcatgccgTGcaacatgtacctgTGttctgcgtta | 790-825 |
| N2 His267/272Arg F | ctgtgatcatgccgcGGaacatgtacctgcGttctgccttagtc | 785-828 |
| N2 Glu299Ala F | acttcttcatcgCgtcctgcatcg | 883-907 |
| N2 Arg416Ala F | gtgatcctgaccGCgtctatcgc | 1234-1256 |
| Primer . | Nucleotide sequence, 5′ to 3′ . | Position in Nramp2 . |
|---|---|---|
| pCB6 F | aatgggcggtaggcgtgta | 848-866 of pCB6 |
| pCB6 R | aggccaggagaggcactgg | 1006-1034 of pCB6 |
| N2 Asp86Ala F | cctacctagCcccaggaaac | 248-267 |
| N2 Arg119Ala F | ctgctgctccagGCccttgcag | 346-367 |
| N2 Arg146Ala F | caaggtcccaGCgatcatcctgtg | 426-449 |
| N2 Glu154Ala F | ctgatggtggCgttggcaatca | 451-472 |
| N2 Asp161Ala F | cattggttctgCcatgcaggaagtc | 471-495 |
| N2 Gly184Asp F | ccctgtgggAcggagtcctca | 542-563 |
| N2 Gly185Arg F | gggtccccctgtggggcaGagtcctcatcaccatc | 536-570 |
| N2 Asp192Ala F | caccatcgcagCcacttttgtg | 564-585 |
| N2 Glu225Ala F | gtttggatatgCgtacattac | 663-684 |
| N2 His267Ala F | atcatgccgGCcaacatgtacct | 790-825 |
| N2 His267Cys F | atcatgccgTGcaacatgtacct | 790-825 |
| N2 His267Arg F | atcatgccgcGGaacatgtacct | 790-825 |
| N2 His272Ala F | aacatgtacctgGCttctgcctta | 802-825 |
| N2 His272Cys F | aacatgtacctgTGttctgcctta | 802-825 |
| N2 His272Arg F | atgtacctgcGttctgcctta | 805-825 |
| N2 His267/272Ala F | atcatgccgGCcaacatgtacctgGCttctgcgtta | 790-825 |
| N2 His267/272Cys F | atcatgccgTGcaacatgtacctgTGttctgcgtta | 790-825 |
| N2 His267/272Arg F | ctgtgatcatgccgcGGaacatgtacctgcGttctgccttagtc | 785-828 |
| N2 Glu299Ala F | acttcttcatcgCgtcctgcatcg | 883-907 |
| N2 Arg416Ala F | gtgatcctgaccGCgtctatcgc | 1234-1256 |
F indicates forward; R, reverse. Uppercase letters in nucleotide sequences indicate bases that differ from theNramp2 sequence.