Table 2 Oligonucleotide primer sequences

539 frag1 5'

ureC upstream

Forward

5'-GACCTTTACCCACAGCTAAT-3'

539 frag1 3'

ureC upstream

Reverse

5'-TAGTTAGTCACTTGAAATTGTTA

ATGCCAT-3'

539 frag2 5'

Kanamycin cassette

Forward

5'-CAATTTCAAGTGACTAACTAGGA

GGAATAA-3'

539frag2 3'

Kanamycin cassette

Reverse

5'-TGACCCAATGCATTATTCCCTCC

AGGTACT-3'

539 frag3 5'

ureC downstream

Forward

5'-GGGAATAATGCATTGGGTCAGC

GATA-3'

539 frag3 3'

ureC downstream

Reverse

5'-ATCGCACACCGAGTTTG-3'

Cloning of ureC into pSPEC1 to complement ureC mutation

539promoter

F1

ureA upstream

Forward

5'-GAGAGGATCCGTAAAATTCGCTG

ACTTTCG-3'

539C R1

ureC downstream

Reverse

5'-ATATGAATTCGCTACTTCACGCC

CCGTA-3'

Construction of urease operon mutant

539 Op frag1 F1

ureA upstream

Forward

5'-TACACCTTCCTTGCCCAC-3'

539 Op frag1 R1

ureA upstream

Reverse

5'-TAGTTAGTCAATTTTCATTCCTTAAT-3'

539 Op frag2 F1

Kanamycin cassette

Forward

5'-GAATGAAAATTGACTAACTAG

GAGGAATAA-3'

539 Op frag2 R1

Kanamycin cassette

Reverse

5'-ACCAATTTTCCATTATTCCCTCCA

GGTACT-3'

539Op frag3 F1

ureA downstream

Forward

5'-GGGAATAATGGAAAATTGGTAG

GCTAT-3'

539Op frag3 R1

ureA downstream

Reverse

5'-CAGATGTTGCTTCAATTAAG-3'

Subjecting multiple strains to PCR to assess presence of urease operon

UreaseA F1

ureA

Forward

5'-ATGCACTTAACTTCCAGAG-3'

UreaseA R1

ureA

Reverse

5'-TTATCTGATTGGATTATGC-3'

539 F1

ureC

Forward

5'-CAACATGGCATTAACAATTTCAAG-3'

539 R1

ureC

Reverse

5'-TTAGAATAGGAAATATCGCTG-3'

UreaseH F1

ureH

Forward

5'-ATGAACAGTAAATTATCC-3'

UreaseHR1

ureH

Reverse

5'-GAATTTGCTCTGCACGACA-3'

Cloning of ureC gene to express recombinant protein

539 F1

ureC

Forward

5'-CAACATGGCATTAACAATTTCAAG-3'

539 R1

ureC

Reverse

5'-TTAGAATAGGAAATATCGCTG-3'

Performing RT PCR

UreA-UreB F1

ureA, ureB

Forward

5'-ATGAGTGTAGCGGAAGTG-3'

UreA-UreB R1

ureA, ureB

Reverse

5'-ACGTTTAAACGCATTCCGC-3'

UreB-UreC F1

ureB, ureC

Forward

5'-TGAAACCAATAATGCCCT-3'

UreB-UreC R1

ureB, ureC

Reverse

5'-TTGCGTGCGTACCATCAGC-3'

UreC-UreE F1

ureC, ureE

Forward

5'-GGTATTGCGGAGCATATTGG-3'

UreC-UreE R1

ureC, ureE

Reverse

5'-GAATGACTGTGAGAATGCC-3'

UreE-UreF F1

ureE, ureF

Forward

5'-GCTTGAAACACGATGATG-3'

UreE-UreF R1

ureE, ureF

Reverse

5'-TGATTGCACCGACTGCTG-3'

UreF-UreG F1

ureF, ureG

Forward

5'-TGCCTTCGATGCAATGAAC-3'

UreF-UreG R1

ureF, ureG

Reverse

5'-AGCACCAACGAATGGAGC-3'

UreG-UreH F1

ureG, ureH

Forward

5'-AGTTTGCTTCCACCAGAGC-3'

UreG-UreH R1

ureG, ureH

Reverse

5'-GTTCGTCATTCAACACCC-3'