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Table I.

A List of GC Laboratory Strains, Oligonucleotide Primers, and Plasmids Used


GC strains

Poly-G in pgtA

Isolated from

Site of isolation

Accession no.

Reference
FA1090  +, (11 G, on) DGI Cervix AF485418 (73) 
15253  +, (14 G, on) DGI Blood AF485418 (74) 
UUI +, ∼ (10 G, off) DGI Blood AF485418 (75) and a 
KH4318 +, ∼ (11 G, on) AF485418 
M20 +, ∼ (11 G, on) AF485418 
RT397 +, ∼ (11 G, on) AF485418 
F62 — UG Female urethra AF485419 (73) 
MS11A/C — UG AF485419 (38) 
1291 — UG Male urethra AF485419 (76) and b 
R10 — UG Male urethra AF485419 (38) 
M13Ison — UG AF485419 c 
140Ison — UG AF485419 c 
Pgh3-2
 

 
UG
 
?
 
AF485419
 
(38)
 
Primers Direction 59 Extension Sequence Position Reference 
PgtA1 CGGCGGAACAGGTTAAG 1,802 bp of ppgtA1 insert Fig. 2 A 
PgtA2 TCGATCAGGAAGCCGTTG 1,999 bp of ppgtA1 insert Fig. 2 A 
PgtA3 ATGATCGGATCC AATTCGGTGACTACGGCTG 684 bp of ppgtA1 insert Fig. 2 A 
PgtA4 ATACCGGAATTC ACCACCTTGTTTACGCCTT 2,279 bp of ppgtA1 insert Fig. 2 A 
PgtA5 ATGTAAGAGCTC AATTCGGTGACTACGGCTG 684 bp of ppgtA1 insert Fig. 2 A 
PgtA6
 
R
 
ATACCGGATATC
 
TCGATCAGGAAGCCGTTG
 
1,999 bp of ppgtA1 insert
 
Fig. 2 A
 
Plasmids Insert description    Reference 
ppgtA1 3,356 bp F62 pgtA insert starting 1,138 bp upstream of start codon    Fig. 2 A 
ppgtA2 MS11A pgtA DNA equivalent to 1,205–2,268 bp of ppgtA1 insert    Fig. 2 A 
ppgtA3 FA1090 pgtA DNA equivalent to 1,445–3,345 bp of ppgtA1 insert    Fig. 2 A 
ppgtA4 15253 pgtA DNA equivalent to 684–2,668 bp of ppgtA1 insert    Fig. 2 A 
ppgtA5 FA1090 PgtA3-PgtA4 amplicon cloned in pBluescript II KS+    Fig. 2 A 
ppgtA5-erm An XmaI-BsaWI ermC′ (24) fragment cloned in AgeI site of ppgtA5 to knockout pgtA    Fig. 2 A 
ppgtA-F1a FA1090 PgtA5-PgtA6 amplicon cloned in pCMVTag4b. In-frame COOH-terminal fusion
 of FLAG epitope tag with the Pv+ (11G) pgtA   Fig. 2 A 
ppgtA-F1b Identical insert as above cloned in pCMVTag4c. Out of frame Pv+ pgtA–FLAG fusion.    Fig. 2 A 
ppgtA-F2a F62 PgtA5-PgtA6 amplicon cloned in pCMVTag4b. In-frame fusion of FLAG with PvpgtA   Fig. 2 A 
ppgtA-F2b
 
Identical insert as above cloned in pCMVTag4a. Out of frame FLAG fusion with the PvpgtA.
 
   Fig. 2 A
 

GC strains

Poly-G in pgtA

Isolated from

Site of isolation

Accession no.

Reference
FA1090  +, (11 G, on) DGI Cervix AF485418 (73) 
15253  +, (14 G, on) DGI Blood AF485418 (74) 
UUI +, ∼ (10 G, off) DGI Blood AF485418 (75) and a 
KH4318 +, ∼ (11 G, on) AF485418 
M20 +, ∼ (11 G, on) AF485418 
RT397 +, ∼ (11 G, on) AF485418 
F62 — UG Female urethra AF485419 (73) 
MS11A/C — UG AF485419 (38) 
1291 — UG Male urethra AF485419 (76) and b 
R10 — UG Male urethra AF485419 (38) 
M13Ison — UG AF485419 c 
140Ison — UG AF485419 c 
Pgh3-2
 

 
UG
 
?
 
AF485419
 
(38)
 
Primers Direction 59 Extension Sequence Position Reference 
PgtA1 CGGCGGAACAGGTTAAG 1,802 bp of ppgtA1 insert Fig. 2 A 
PgtA2 TCGATCAGGAAGCCGTTG 1,999 bp of ppgtA1 insert Fig. 2 A 
PgtA3 ATGATCGGATCC AATTCGGTGACTACGGCTG 684 bp of ppgtA1 insert Fig. 2 A 
PgtA4 ATACCGGAATTC ACCACCTTGTTTACGCCTT 2,279 bp of ppgtA1 insert Fig. 2 A 
PgtA5 ATGTAAGAGCTC AATTCGGTGACTACGGCTG 684 bp of ppgtA1 insert Fig. 2 A 
PgtA6
 
R
 
ATACCGGATATC
 
TCGATCAGGAAGCCGTTG
 
1,999 bp of ppgtA1 insert
 
Fig. 2 A
 
Plasmids Insert description    Reference 
ppgtA1 3,356 bp F62 pgtA insert starting 1,138 bp upstream of start codon    Fig. 2 A 
ppgtA2 MS11A pgtA DNA equivalent to 1,205–2,268 bp of ppgtA1 insert    Fig. 2 A 
ppgtA3 FA1090 pgtA DNA equivalent to 1,445–3,345 bp of ppgtA1 insert    Fig. 2 A 
ppgtA4 15253 pgtA DNA equivalent to 684–2,668 bp of ppgtA1 insert    Fig. 2 A 
ppgtA5 FA1090 PgtA3-PgtA4 amplicon cloned in pBluescript II KS+    Fig. 2 A 
ppgtA5-erm An XmaI-BsaWI ermC′ (24) fragment cloned in AgeI site of ppgtA5 to knockout pgtA    Fig. 2 A 
ppgtA-F1a FA1090 PgtA5-PgtA6 amplicon cloned in pCMVTag4b. In-frame COOH-terminal fusion
 of FLAG epitope tag with the Pv+ (11G) pgtA   Fig. 2 A 
ppgtA-F1b Identical insert as above cloned in pCMVTag4c. Out of frame Pv+ pgtA–FLAG fusion.    Fig. 2 A 
ppgtA-F2a F62 PgtA5-PgtA6 amplicon cloned in pCMVTag4b. In-frame fusion of FLAG with PvpgtA   Fig. 2 A 
ppgtA-F2b
 
Identical insert as above cloned in pCMVTag4a. Out of frame FLAG fusion with the PvpgtA.
 
   Fig. 2 A
 
a

Z. McGee,

b

M. Apicella, and

c

C. Ison, personal communications.

Restriction sites in the primers are underlined. ppgtA1, ppgtA2, ppgtA3, and ppgtA4 are from GC libraries made by cloning partially digested fragments of genomic Tsp509I in pBluescript II SK. Except in ppgtA1, DNA inserts are in the direction of lacZ in these plasmids. +, present; −, absent; ?, unknown; ∼, approximate data; DGI, disseminated gonococcal infection; UG, uncomplicated gonorrhea; Pv+, phase-variable pgtA; Pv, pgtA that expresses constitutively without any variation; F, forward or in the direction of pgtA; R, direction reverse to F. The number of Gs in each poly-G and expression status of pgtA are indicated in parentheses.

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