Table 4.
Oligonucleotides
| Primer | Sequence (5’3’)a, b | Use/locus tag/reference |
|---|---|---|
| oMC305 | CACAGGAGGTATCGTACAG | Forward primer for sequencing Spo0A |
| oMC306 | GCGAAACGGTATAACCCTAG | Reverse for sequencing Spo0A |
| oMC1249 | GTCGAGGATCCGATGACAAGTTATTGGAATACACAG | Forward primer for Spo0A expression from pMC123 |
| oMC1250 | GACTCGAATTCCCCTAGTGGTTATACCGTTTCG | Reverse primer for Spo0A expression from pMC123 |
| oMC1251 | ATTAATACTAGCTGTAATAATGCCACATC | Forward SOEing primer for Spo0A D56A |
| oMC1252 | GAT GTGGCATTATTACAGCTAGTATTAAT | Reverse SOEing primer for Spo0A D56A |
| oMC1513 | GTTTTAGCAGATGACGCTAAGGATTTTTGTCAG | Forward SOEing primer for Spo0A N12A |
| oMC1514 | CTGACAAAAATCCTTAGCGTCATCTGCTAAAAC | Reverse SOEing primer for Spo0A N12A |
| oMC1515 | TTAGCAGATGACAATGCAGATTTTTGTCAGGTA | Forward SOEing primer for Spo0A K13A |
| oMC1516 | TACCTGACAAAAATCTGCATTGTCATCTGCTAA | Reverse SOEing primer for Spo0A K13A |
| oMC1517 | AAGGATTTTTGTCAGGCATTAAAAGAGTATTTG | Forward SOEing primer for Spo0A V18A |
| oMC1518 | CAAATACTCTTTTAATGCCTGACAAAAATCCTT | Reverse SOEing primer for Spo0A V18A |
| oMC1519 | TTAATACTAGATGTAGCAATGCCACATCTAGAT | Forward SOEing primer for Spo0A I58A |
| oMC1520 | ATCTAGATGTGGCATTGCTACATCTAGTATTAA | Reverse SOEing primer for Spo0A I58A |
| oMC1547 | GATGCGAATTCTCACTTGTCATCGTCATCCTTGTAA TCTATGTCATGATCTTTATAATCACCGTCATGGT CTTTGTAGTCACCTCCTTTAACCATACTATGTTC TAGTCTTAA |
Reverse primer for Spo0A with 3x FLAG tag and homology to pMC123 |
| oMC1583 | GATGTAATAATGCCAGCACTAGATGGATTAGGT | Forward SOEing primer for Spo0A H61A |
| oMC1584 | ACCTAATCCATCTAGTGCTGGCATTATTACATC | Reverse SOEing primer for Spo0A H61A |
| oMC2354 | AGGTTATAGACTTTTTGAAGAAATTCTATAGCT CGATCGGTGTAAAAAGTTTAGTTTTCTGTAATA AGAAGATGT |
Forward primer to amplify Spo0A fused to LgBit fragment |
| oMC2437 | CTTGATCGTAGCGTTAACAGATCTGAGCTCGTG TAAAAAGTTTAGTTTTCTGTAATAAGAAGATGT |
Forward primer to amplify Spo0A fused to SmBit fragment |
| oMC2439 | CACCACCACTAGAACCCCCTCGAGATTTAACCAT ACTATGTTCTAGTCTTAATTTATCAGC |
Reverse primer to amplify Spo0A fused to SmBit fragment |
| oMC2447 | ACCACCACCACTAGAACCTGCGGCCGCTCCTTTAA CCATACTATGTTCTAGTCTTAATTTATCAGC |
Reverse primer to amplify Spo0A fused to LgBit fragment |
a
Restriction sites underlined
b
Nucleotides for site-directed mutagenesis are noted in bold