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. 2021 Dec 4;10(1):2205–2219. doi: 10.1080/22221751.2021.2002671

Table 1.

Resistant strains and mutations conveying phage resistance and outcome of wildtype gene complementations in trans.

Resistant Isolate Mutation Putative gene function Complementation
Yes No
R1 abcT (A1844T, p.Glu615Val) ABC transporter   abcT
R1 actP (A1144G, p.Thr382Ala) acetate permease   actP
R1 gtrOC3 (IS4 family insertion) LPS/ LOS biosynthesis gtrOC3  
R1 phoH (1011nt, 103th A loss) phosphohydrolase   phoH
R1 decT (753nt, G233A, p.Thr78Met) di-trans,poly-cis-decaprenylcistransferase    
R1 dcaP (1332nt, G395T, p.Pro132Glu) DcaP-like protein    
R1 Udp (720nt, T35C, p.Leu 12 Ser) UDP-2,3-diacylglucosamine diphosphatase    
R2 abcT (A1844T, p.Glu615Val) ABC transporter   abcT
R2 actP (A1144G, p.Thr382Ala) acetate permease   actP
R2 gtrOC3 (535th A loss, p.Asn179Ile fs Ter7) LPS/ LOS biosynthesis gtrOC3  
R2 phoH (1011nt, 103th A loss) phosphohydrolase   phoH
R2 decT (753nt, G233A, p.Thr78Met) di-trans,poly-cis-decaprenylcistransferase    
R2 dcaP (1332nt, G395T, p.Pro132Glu) DcaP-like protein    
R2 Udp (720nt, T35C, p.Leu 12 Ser) UDP-2,3-diacylglucosamine diphosphatase    
R5 gtr9 (IS4 insertion) Capsule biosynthesis   gtr9
R5 abcT (A1844T, p.Glu615Val) ABC transporter   abcT
R5 actP (A1144G, p.Thr382Ala) acetate permease   actP
R5 gtr0C3 (535th A loss, p.Asn179Ile fs Ter7) LPS/ LOS biosynthesis   gtr0C3
R5 phoH (1011nt, 103th A loss) phosphohydrolase   phoH
R5 decT (753nt, G233A, p.Thr78Met) di-trans,poly-cis-decaprenylcistransferase    
R5 dcaP (1332nt, G395T, p.Pro132Glu) DcaP-like protein    
R5 Udp (720nt, T35C, p.Leu 12 Ser) UDP-2,3-diacylglucosamine diphosphatase    
R6 abcT (A1844T, p.Glu615Val) ABC transporter   abcT
R6 actP (A1144G, p.Thr382Ala) an acetate permease   actP
R6 phoH (1011nt, 103th A loss) phosphohydrolase   phoH
R6 decT (753nt, G233A, p.Thr78Met) di-trans,poly-cis-decaprenylcistransferase    
R6 dcaP (1332nt, G395T, p.Pro132Glu) DcaP-like protein    
R6 Udp (720nt, T35C, p.Leu 12 Ser) UDP-2,3-diacylglucosamine diphosphatase    
R7 gtr9 (IS5 insertion) Capsule biosynthesis gtr9  
R7 abcT (A1844T, p.Glu615Val) ABC transporter   abcT
R7 actP (A1144G, p.Thr382Ala) an acetate permease   actP
R7 phoH (1011nt, 103th A loss) phosphohydrolase   phoH
R7 decT (753nt, G233A, p.Thr78Met) di-trans,poly-cis-decaprenylcistransferase    
R7 dcaP (1332nt, G395T, p.Pro132Glu) DcaP-like protein    
R7 Udp (720nt, T35C, p.Leu 12 Ser) UDP-2,3-diacylglucosamine    
R8 abcT (A1844T, p.Glu615Val) ABC transporter   abcT
R8 actP (A1144G, p.Thr382Ala) an acetate permease   actP
R8 gtrOC3 (535th A loss, p.Asn179Ile fs Ter7) LPS/ LOS biosynthesis gtrOC3  
R8 phoH (1011nt, 103th A loss) phosphohydrolase   phoH
R8 decT (753nt, G233A, p.Thr78Met) di-trans,poly-cis-decaprenylcistransferase    
R8 dcaP (1332nt, G395T, p.Pro132Glu) DcaP-like protein    
R8 Udp (720nt, T35C, p.Leu 12 Ser) UDP-2,3-diacylglucosamine diphosphatase    
R10 gtr9 (559th T loss, p. Leu187Tyr fs Ter3) Capsule biosynthesis gtr9  
R10 abcT (A1844T, p.Glu615Val) ABC transporter   abcT
R10 actP (A1144G, p.Thr382Ala) an acetate permease   actP
R10 phoH (1011nt, 103th A loss) phosphohydrolase    
R10 decT (753nt, G233A, p.Thr78Met) di-trans,poly-cis-decaprenylcistransferase    
R10 dcaP (1332nt, G395T, p.Pro132Glu) DcaP-like protein    
R10 Udp (720nt, T35C, p.Leu 12 Ser) UDP-2,3-diacylglucosamine diphosphatase    
R10 isoS (1698nt, 655th T loss, p.Ser 219 Pro fs Ter23) 2-isopropylmalate synthase    
R10 LptD (2439nt, 2395th G to T, p.Val 799 Phe) LPS- assembly protein    
R10 long-chain fatty acid--CoA ligase (1680nt,1240th T to A, p.Phe 414 Ile) long-chain fatty acid--CoA ligase    
R12 abcT (1A1844T, p.Glu615Val) ABC transporter   abcT
R12 actP (A1144G, p.Thr382Ala) an acetate permease   actP
R12 gtrOC3 (535th A loss, p.Asn179Ile fs Ter7) LPS/ LOS biosynthesis gtrOC3  
R12 phoH (1011nt, 103th A loss) phosphohydrolase   phoH
R12 decT (753nt, G233A, p.Thr78Met) di-trans,poly-cis-decaprenylcistransferase    
R12 dcaP (1332nt, G395T, p.Pro132Glu) DcaP-like protein    
R12 Udp (720nt, T35C, p.Leu 12 Ser) UDP-2,3-diacylglucosamine diphosphatase    
R13 No PCR product of gtr9 Capsule biosynthesis   gtr9
R13 abcT (A1844T, p.Glu615Val) ABC transporter   abcT
R13 actP (A1144G, p.Thr382Ala) an acetate permease   actP
R13 phoH (1011nt, 103th A loss) phosphohydrolase   phoH
R13 decT (753nt, G233A, p.Thr78Met) di-trans,poly-cis-decaprenylcistransferase    
R13 dcaP (1332nt, G395T, p.Pro132Glu) DcaP-like protein    
R13 Udp (720nt, T35C, p.Leu 12 Ser) UDP-2,3-diacylglucosamine diphosphatase    
R28 gtr9 (transposase insertion) Capsule biosynthesis   gtr9
R33 gtr9 (IS5 insertion) Capsule biosynthesis   gtr9
R35 gtrOC3 (535th A loss, p.Asn179Ile fs Ter7) LPS/ LOS biosynthesis gtrOC3  
R39 gtrOC3 (535th A loss, p.Asn179Ile fs Ter7) LPS/ LOS biosynthesis gtrOC3  
R54 gtr9 (transposase insertion) Capsule biosynthesis   gtr9
R78 gtrOC3 (IS5 family insertion) LPS/ LOS biosynthesis gtrOC3  
gtr9 KO Markerless knock out the entire gtr9 on the background of ATCC17978 Capsule biosynthesis gtr9  
gtrOC3-delta A Subsititute for the wild type gtr0C3 on the background of ATCC17978 LPS/ LOS biosynthesis gtrOC3  
R81 gtr9 (transposase insertion) Capsule biosynthesis gtr9  
R83 No PCR product of gtr9 Capsule biosynthesis   gtr9
R86 gtr9 (transposase insertion) Capsule biosynthesis gtr9  
R115 gtr9 (transposase insertion) Capsule biosynthesis gtr9  
R125 gtr1 (448th A loss, p. Met 150 Cys fs Ter22) Glycosyltransferase   gtr1
R130 gtr9 (transposase insertion) Capsule biosynthesis gtr9  
R132 gtr9 (transposase insertion) Capsule biosynthesis   gtr9
R132 gtr1 (448th A loss, p. Met 150 Cys fs Ter22) Glycosyltransferase   gtr1
R134 gtr9 (transposase insertion) Capsule biosynthesis gtr9  
XH198 gtr9 KO Markerless knock out the entire gtr9 on the background of XH198 Capsule biosynthesis gtr9  
XH198 gtrOC3-KO Markerless knock out the entire gtrOC3 on the background of XH198 LPS/ LOS biosynthesis gtrOC3  
R518 gtrOC3 (535th A loss, p.Asn179Ile fs Ter7) LPS/ LOS biosynthesis gtrOC3  
R587 gtrOC3 (535th A loss, p.Asn179Ile fs Ter7) LPS/ LOS biosynthesis gtrOC3  

Yes: After the transformation of the corresponding plasmid which contains the WT gene, the R variant can get infected by Phab24.

No: After the transformation of the corresponding plasmid which contains the WT gene, the R variant cannot get infected by Phab24.

R1-R80 are Phab24 escape mutants from ATCC17978.

R81-R587 are Phab24 escape variants from XH198.

Based on the NCBI accession number CP018664.1, ATCC17978:

AUO97_06900, gtr9, 828nt

AUO97_07355, abcT, 1932nt

AUO97_05550, actP, 1716nt

AUO97_03485, gtrOC3, 768nt

AUO97_03925, phoH, 1011nt

AUO97_06920, gtr1, 1164nt

AUO97_09800, 2-isopropylmalate synthase, isoS, 1698nt

AUO97_15295, LPS assembly protein LptD, 2439nt

AUO97_06215, long-chain fatty acid--CoA ligase, 1680nt

AUO97_17750, di-trans, poly-cis-decaprenylcistransferase, decT, 753nt

AUO97_18665, UDP-2,3-diacylglucosamine, Udp, 720nt

Interestingly, the phoH of the WT or ATCC17978 used for Sanger sequencing has 1012nt, while the phoH of CP018664.1 has 1011nt, the same as the mutated R variants. Maybe that is the reason why the phoH is annotated as the pseudogene.