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PF8 Tel + (33) 1 45 68 83 24 |
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- Genes The Kingella kingae MLST scheme uses internal fragments of the following six housekeeping genes:
- Primers for PCR amplification
abcZ : abcZ-F : TGACGACCAAGCGAGCGTGTTTGA
abcZ : abcZ-R : TCCGCCTCAACCGCCAATTCCT
adk : adk-F : CACAAGCGCAATTTATTACGCGCGA
adk : adk-R : GTCGTCATCGCGTTGCACCAAGT
aroE : aroE-F : TTTGCCGCACAAGAAGGTGCGCAA
aroE : aroE-R : GGCTGGTTCGCATAAAACATATCG
cpn60 : cpn60-F : TGTTGGCGCAAGCGATTGTTGCT
cpn60 : cpn60-F-3 : AAACCAATGATGTGGCTGGCGACG (For few strains, we use this forward primer in case of failure with the above primer)
cpn60 : cpn60-R : AATGGGCTGTCCAAACCAGCGAT
gdh : gdh-F : CGGCGCGTTGCGCGATATGGTGCA
gdh : gdh-R : CCAGTTGTCCAAAATCGGCATCAC
recA : recA-F : GACGAAGAATTGCAAGTCATTTCCA
recA : recA-R : AGTTTACGCAAGGCTTGGCTCATC
- Sequencing primers
The same primers as for PCR are used.
- PCR conditions Conditions for housekeeping genes amplification were 15 minutes at 95°C followed by 35 cycles of 95°C for 15 s, 56°C for 30 s and 72°C for 1 min 30 s with a final extension step at 72°C for 10 minutes. - Allele templates (K. kingae ATCC23330 alleles) abcZ (453 bp)
TTGGGCGATTTGCGCGATGTGTTGCGCCGTTATCATCAGGTTAGCCACGCGCTAGAAACGCAGCATTCTGATGATTTAATTCGCCA
ACTCAATGAGTTACAAAATGAAATTGAATCGCAAGACGGCTGGCAGTTTGACGCGGCAATCCGCCAAACGATTAGCGAATTGGGGC TGCCTGAAAATGAATTGATTGGCAACTTGTCGGGTGGTCAGAAAAAACGGGTTGCGTTGGCTCAAGCGTGGGTGCAAAAACCCGAT ATTTTGTTGCTGGACGAACCGACTAACCATTTGGATATTAACGCGATTATTTGGCTGGAAAATTTGTTGCAAGCGTTTTCAGGCAG CATGGTGGTGATTACGCACGACCGCCGTTTTTTGGACAACATCGCCAATCGGATTGTGGAATTGGATAGGGGCGTGTTGCGTTCGT ACGAGGGCAGTTTCAGCAAATAC adk (351 bp)
ATTTCTACTGGCGATATGCTTCGCGCGGCGATTAAAGCTGGCACACCATTGGGTTTGGAAGCCAAAAAAATCATGGACGAAGGCG
GCTTGGTGTGCGATGACATCATCATCGGCATGGTCAAAGAGCGCGTGGCACAGCCCGATTGCGCCAATGGTTTTTTGTTTGATGG CTTTCCACGCACCTTGGCACAAGCCGAAGCGATGATTGAAGCAGGTGTGCATTTGGACGCTGTGGTCGAAATCGACGTACCCGAC GAAACAATCGTAGAACGCATGAGCGGTCGCCGTGTGCATTTGGCATCGGGTCGCACTTATCACTTGAAGCACAATCCGCCCAAAG TGGCTGGCAAA aroE (519 bp)
CGCACCTTTGCCGAAAACACCGCACAAGATTTTGGCGATGGCGTTCGCGCTTTTTTTGCACAAAATGGACGTGGAGCAAATGTGA
CCCTGCCGTTTAAAGGTTTTGCCTACGATTTGGTGCAGGCACACACCGAACGCGCTCGGGCAGCGGGCGCAGTCAATACGCTGAT TCCGCAAGCAGACGGCACTTTGCTGGGCGACAACACCGACGGCGAAGGCTTGGTGCGCGACTTATGCCAGCACTTGCAATTCAAT CTACACAATAAAAACATTTTGCTTTTGGGCGCAGGTGGCGCGACTCGTGGCGTGGTTTTGCCGCTAGCAGCCTGCAATCCTGCCA GCATCACCATTGCCAATCGCACACACGATAAAGCCATTGAATTGGCTCAGTTGTTTGATATTGCCGCTCTGCCCTTTCATCAATT AAATCAATCTTATGACATTATTATCAACGCAACATCGGGTAGCATAGACGATAGCATTCCAGATGTATCGCCCGATATTTTTGCA GGCTGCTCG cpn60 (303 bp)
GTTACCGCAGGCATGAATCCTACCGATTTGAAACGCGGTATTGACAAAGCAGTGGCGGCTTTGGTTGGCGAATTGGCAAACATCG
CGAAACCTTGCGAAACATACGAGCAAATCGCTCAAGTGGGCGCGATTTCTGCGAACTCTGACGAGCAAGTTGGCAAAATCATTGC AGACGCGATGCAAGAAGTCGGCAAAGAGGGCGTGATTACCGTTGAAGACGGCAAATCATTGGAAAACGAGTTAGAAGTGGTTAAA GGTATGCAATTTGACCGTGGCTACTTGTCGCCTTATTTCGTGAATGAT gdh (576 bp)
ATGCAAATGCTGTGTTTTGTGGCAATGGAACAACCCAAATCGTTGAGCGCAGACGATGTACGCGATGAAAAATTGAAAGTGGTGG
CTGCACTCAAACCGATGACTTCGGCAGATGTAGATAAAAACGTGATTCGCGCACAATATGTGGCAAACGGCGAGCAAAACGGCTA TTTGCAAGAACACAATGTGCCAGCCGACAGCCGTACCGAAACCTATGTTGCCATTCGCGCCGAAATTGATACGCCACGTTGGGCA GGCGTGCCGTTTTATTTGCGTACAGGCAAACGCTTGGCAACGCGCACAGCAGAAATTGTTTTGAATTTCAAAGACCAAGCCAATG GTTTGTTTGGCACAAATCCTAATCGTTTGGTGATTTCGTTGCAGCCCGATGAAACCGTATCGCTCAAATTGTATGTGAAACAAGT GGGCAGCGGCTTGGACGTGCAGCCAACCGAAATGGTGTTGGACATGGGCACAGTGTCAGACGAACGCCGCGCCGAAGCGTATGAA TTGCTGTTGCGCGAAGTGATTGACGGACGTTTGAGCTTGTTCAATCGCCGCGACGAGCTGGAAAAA recA (354 bp)
TTGGACATTGCCTTGGGTGTGGGCGGTTTACCACGTGGTCGTATTGTGGAAATTTACGGCGCAGAATCATCGGGCAAAACCACTT
TATGTTTGGAATCGATTGCACAATGCCAAAAAGCAGGCGGAACGTGTGCGTTTATTGACGCAGAACACGCATTTGACCCTATTTA CGCGCGTAAATTGGGCGTAGAAGTCGAAAAATTGTTGGTATCGCAACCCGACACAGGCGAACAAGCTTTGGAAATTTGCGATATG TTGGTGCGCTCGGGCGGTGTGGATATGGTCGTGATTGATTCGGTTGCCGCACTTGTTCCCAAAGCGGAAATTGAAGGCGACATGG GCGACAGCCACGTT |
