Table 1.
PCR primers
| Primer specifity | Primer symbol | Position of bp in cDNA | Sequence |
|---|---|---|---|
| α subunit primers common to all isoforms | CA-1 | 2,086–2,108 | CAGCAGAAGCTCATCATTGTGGA |
| CA-2 | 2,853–2,831 | GTTCTTCATGCCCTGCTGGAAGA | |
| CA-3 | 1,171–1,193 | ATGTGGTTCGACAACCAAATCCA | |
| CA-4 | 1,817–1,795 | CATTTGCCCACAGCATCTGGAAC | |
| α1 subunit isoform-specific primers | A1-1 | 1–26 | ATGGGGAAGGGGGTTGGACGTGATAA |
| A1-2 | 530–503 | TTCTCACCATTTCGAATCACAAGGGCTT | |
| A1-3 | 283–315 | CAGCTCTTTGGGGGGTTCTCAATGTTACTGTGG | |
| A1-4 | 980–948 | ATACCGATGAGGAAGATGACAGCCACAAGCCAG | |
| A1-5 | 628–650 | GGCTGCAAGGTGGATAACTCATC | |
| A1-6 | 1,467–1,444 | CTGGTACTTGTTGGTGGAGTTGAA | |
| A1-7 | 1,751–1,784 | GCTTTGTTGGGCTCATCTCCATGATTGACCCTCC | |
| A1-8 | 2,726–2,693 | TCATAGGTCCACTGCTGCCCGTAGCTGTCTTCCA | |
| A1-9 | 2,548–2,581 | ATCAGCATGGCCTATGGGCAGATTGGAATGATCC | |
| A1-10 | 3,072–3,046 | CTAATAGTAGGTTTCCTTCTCCACCCA | |
| α2 subunit isoform-specific primers | A2-1 | 58–81 | GGCGGCAAGAAGAAACAGAAGGAG |
| A2-2 | 668–645 | TGGGGCTCCGACTCTCCTGTTAAG | |
| A2-3 | 828–852 | GGTTCGGCGGACACCCATAGCAATG | |
| A2-4 | 1,502–1,478 | ACGTGGCTCTGGGGGCTGTCTTCTC | |
| α3 subunit isoform-specific primers | A3-1 | 1–33 | ATGGAGATACACGACAAGAAAGATGACAAGGAC |
| A3-2 | 540–508 | CCCGACCACCACCTCCTCAGCGTTCACCTGCAT | |
| A3-3 | 6–31 | GATACACGACAAGAAAGATGACAAGG | |
| A3-4 | 650–627 | TGGGGCTCGGATTCTCCAGTCAGG | |
| A3-7 | 828–856 | CGCCATCGAGATTGAGCACTTCATCCAGC | |
| A3-8 | 1,224–1,195 | GTGCGAACTCTTGTCAAATGAGGTCCCTGA | |
| A3-11 | 1,459–1,488 | ACCGAGGACCCCAACGACAACCGATACCTG | |
| A3-12 | 2,034–2007 | GGTGTGATTCTCCAGGATCTCGTCGATT | |
| A3-13 | 1,659–1,689 | CGAGGAGCAGTATCCCCAAGGCTTTGCCTTC | |
| A3-14 | 2,067–2,040 | CTGCTGGGGGGATGTGCGGGCGAAGACG | |
| A3-15 | 2,061–2,084 | CAGCAGAAGCTCATCATTGTGGA | |
| A3-16 | 3,048–3,015 | TCAGTAGTAAGTTTCCTTCTCCACCCAACCCCCT | |
| α4 subunit isoform-specific primers | A4-1 | 137–158 | TAACCTTGGAAGAGCTGAGCA |
| A4-2 | 630–611 | TCCTTGTGCAGAGATAAGCC | |
| β1 subunit isoform-specific primers | B1-1 | 1–33 | ATGGCCCGCGGGAAAGCCAAGGAGGAGGGCAGC |
| B1-2 | 441–409 | CTTTCGCTCTCCTCGTTCATGATTAAAGTCTCC | |
| B1-3 | 105–132 | CCTTCTATTCTACGTAATATTTTATGGC | |
| B1-4 | 756–730 | GGGCTGCAGGAGTTTGCCATAGTACGG | |
| B1-5 | 912–882 | TCAGCTCTTAACTTCAATTTTTACATCAAAA | |
| C-B1 | 198–218 | CACATATCAGGACCGAGTGGC | |
| C-B2 | 659–639 | TCTTCATCTCGCTTGCCAGTG | |
| β2 subunit isoform-specific primers | B2-1 | 419–448 | ATGGAGTCCTCAACTACCCCAAACTGGCCT |
| B2-2 | 873–841 | TCAGGTTTTGTTGATGCGGAGTTTGAAGGCCAC | |
| β3 subunit isoform-specific primers | B3-1 | 16–42 | AAGAAGTCCCTCAACCAGAGCCTGGC |
| B3-2 | 429–405 | TGCAACATAAACTGGACCCTTCTGT | |
| γ subunit isoform-specific primers | γF | 1–33 | ATGGACGGTGGCGGCAGCCCCAAGGGGGACGTG |
| γE | 186–154 | TTACGGCTCATCTTCATTGATTTGCCTGCGCTT |
The sequences of isoforms of the Na,K-ATPase α subunit were aligned, and areas of very high or very low homology were selected. Primers were designed to anneal either to highly conserved areas (subunit primers common to all isoforms) or to areas that show minimal homology (isoform-specific primers). The second column of this table shows the name of the primer, the numbering in the third column refers to the coding region of the cDNA of the respective subunit, and the fourth column shows the oligonucleotide sequence (in the 5′ to 3′ orientation). In cases of the α common primers and the primers for α4, for which only part of the human sequence is known (5), the numbering refers to the α1 subunit. Some of the primers were constructed with one base mismatch to the respective sequence to avoid the formation of an internal hairpin loop.