Table 1

Primer used for the generation of OppA mutants

oppA

clone

deletion/mutation (AA)

name

primer sequence (5'-3')

annealing (°C)


ΔCS1

Δ176-243

OppA start

5'-GTGGCGGCCGCGCCTGCAGTTTTTTAG-3'

60°C

CS1 down

5'-TCTTGATTCAACGTTCTTGTCACCT-3'

60°C

CS1 up

5'- AAGAACGTTGAATCAAGAGAACTAGATGAAGC-3'

62°C

OppA end

5'-GGTCCATGGTGGGTACCAAAATAGACCCGGCATATGTAAAA-3'

62°C


ΔCS2

Δ365-372

OppA start

5'-GTGGCGGCCGCGCCTGCAGTTTTTTAG-3'

50°C

CS2 down

5'-TGAGACGTCTGTAAGCTATCTTTATCCATTGAA-3'

50°C

CS2 up

5'-AAAGATAGCTTACAATACGCTAAATCTACATTG-3'

62°C

OppA end

5'-GGTCCATGGTGGGTACCAAAATAGACCCGGCATATGTAAAA-3'

62°C


ΔDC10

Δ366-381

OppA start

5'-GTGGCGGCCGCGCCTGCAGTTTTTTAG-3'

58°C

DC10 down

5'-CTGACCAATTTTGTATTGTAAGCTATCT-3'

58°C

DC10 up

5'-TACAAAATTGGTCAGAAAGGTATAGAAAAC-3'

58°C

OppA end

5'-GGTCCATGGTGGGTACCAAAATAGACCCGGCATATGTAAAA-3'

58°C


ΔCS3

Δ647-675

OppA start

5'-GTGGCGGCCGCGCCTGCAGTTTTTTAG-3'

61°C

CS3 down

5'-GTACAGCTGTGGAGCATTTAAATATCT-3'

61°C

CS3 up

5'-GCTCCACAGCTGTACGATCCAAACTTCAA-3'

60°C

OppA end

5'-GGTCCATGGTGGGTACCAAAATAGACCCGGCATATGTAAAA-3

60°C


ΔWB

Δ712-727

OppA start

5'-GTGGCGGCCGCGCCTGCAGTTTTTTAG-3'

50°C

DC10 down

5'-ATATGCGTTGAAGTTTGGAT-3'

50°C

DC10 up

5'-TATAACGGTGTTGCTAGCACATAC-3'

58°C

OppA end

5'-GGGTCCATGGTGGGTACCAAAATAGACCCGGCATATGTAAAA-3'

58°C


WA3

874GKDSSGKS-GLQSYGKT881

OppA start

5'-GTGGCGGCCGCGCCTGCAGTTTTTTAG-3'

60°C

DC10 down

5'-TACAGATCTGTTGGTTCTATAGTTTTTCCATAACTCTGCAATCCAAAATC-3'

60°C

DC10 up

5'-CAACAGATCTGTATCAGTGGTCTGCAAT-3'

60°C

OppA end

5'-GGGTCCATGGTGGGTACCAAAATAGACCCGGCATATGTAAAA-3'

60°C


Hopfe et al. BMC Microbiology 2011 11:185   doi:10.1186/1471-2180-11-185

Open Data