Skip to main content

Table 1 Oligonucleotide sequences.

From: Development of a self-replicating plasmid system for Mycoplasma hyopneumoniae

PCR product

Size (bp)

Restriction site

Oligonucleotide sequence (5′ - 3′)

M. hyopneumoniae strain 232 oriC- i

2130

Nco I

F - AGGCCATGG TTGTTAATTATTGCTTGAAATTC

  

Spe I

R - AGTACTAGT AAACTTTATAGGAAAGTTCG

M. hyopneumoniae strain 232 oriC- ii

1823

Nco I

F - GATGCCATGG TTGGTTGATAAGTTCGCTTC

  

Spe I

R - GTAGACTAGT CATTCTTTCAATTTGTCATTC

M. hyopneumoniae strain 232 oriC- iii

859

Nco I

F - GATGCCATGG TTGGTTGATAAGTTCGCTTC

  

Spe I

R - GCTCACTAGT GACAATCAGTTTAAAAATTCAC

M. hyopneumoniae strain 232 oriC- iii

859

Aat II

F - GCAGGACGTC TTGGTTGATAAGTTCGCTCC

  

Nco I

R - GACGCCATGG GACAATCAGTTTAAAAATTCAC

M. hyopneumoniae strain 232 oriC- iv

422

Nco I

F - GCATCCATGG ATATTGTGCATGCCCGCGGATAT

  

Spe I

R - GTAGACTAGT CATTCTTTCAATTTGTCATTC

M. hyopneumoniae strain 232 oriC- v

280

Aat II

F - GCAGGACGTC TTGGTTGATAAGTTCGCTCC

  

Nco I

R - CGTACCATGG CCTAATTTGCTGTGTTCTAA

tetM + spiralin promoter from plasmid pSRT2

2320

Pst I

F - TAACTGCAG CAAAAGCTTGCATGCCTGCA

  

Spe I

R - GTCACTAGT CAGTTCAGATCTTTATATAAC

tetM from plasmid pIVT

2109

Spe I

F - ATCTACTAGT CATGTGATTCTAAAGTATCC

  

Pst I

R - GATCTGCAG CTTATTTAACAAGAAACC

tetM (without promoter) from plasmid pSRT2

1963

Bam HI/Pst I

F - GAACTGCAGGATCC ATGGAGGAAAATCACATGA

  

Sal I

R - CCCGGTCGACTTATATAACAACTTAAATTAC

M. hyopneumoniae strain 232 P97 promoter

619

Bam HI

F - GAATGGATCCCCAACAATTCCGGCAGTC

  

Spe I

R - GCTAACTAGT ACGGGGATTTAAAACAGAAAC

M. hyopneumoniae strain 232 ldh promoter

421

Spe I

F - GGCTAACTAGT ATAGAATTTTGCAATTAAAG

  

Bam HI

R - CTTAGGATCC ATTTATACTCGTATTTGTTAT

M. hyorhinis oriC

1479

Nco I

F - GACTCCATGG TCGGAATCGAGATTCCTAATT

  

Spe I

R - GCCTACTAGT CTACACCTTCGATTTCTCTAA

M. pulmonis oriC

1909

Nco I

F - ATTAGCCATGG CACTCTGGTCAGCGCTAGATC

  

Nhe I

R - TCGCGCTAGC CTAACTTGAAAATAAGCTCC

M. conjunctivae oriC

2307

Nco I

F - GATACCATGG TATGCCTTTGGTCTAGGATG

  

Xba I

R - CGATGTCTAGA CCCTTGAACAAATATACCA

M. ovipneumoniae oriC

2102

Nco I

F – AGTCCATGGTCAATTATTGCTTGGAATTC

  

Spe I

R - CGTACTAGTAAACTTTATAAGAAAGTTCGCT

bla gene (DIG-labelled probe)

860

N/A

F – CCAATGCTTAATCAGTGAGG

   

R - GTATGAGTATTCAACATTTCCG

tetM (DIG-labelled probe)

406

N/A

F – GTGGACAAAGGTACAACGAG

   

R - CGGTAAAGTTCGTCACACAC

  1. Oligonucleotide sequences are shown along with the size of the PCR product generated and the template DNA used. Restriction enzyme sites (where applicable) that were added to the 5′ end of the oligonucleotide are shown in italics. Forward (F) and reverse (R) oligonucleotides are shown.