Strains |
|
|
A. hydrophila SSU |
|
CDC, Atlanta, GA |
SSU-R |
Rifr strain of A. hydrophila SSU |
Laboratory stock |
WT/pBReno |
WT A. hydrophila SSU-R transformed with pBReno plasmid; Rifr Tcr
|
This study |
Enolase mutant |
The chromosomal enolase gene was deleted from WT A. hydrophila SSU-R that was transformed with the plasmid pBReno; Sm/Spr Rifr Tcr
|
This study |
E. coli
|
|
|
SM10 |
Kmr λpir |
24 |
JM109 |
endA1 recA1 gyrA96 thi-1 hsdR17(rK− mK+) relA1 supE44 Δ(lac-proAB) [F′ traD36 proAB lacIqZΔM15] |
Promega |
ES1301 mutS
|
lacZ53 mutS201::Tn5 thyA36 rha-5 metB1 deoC
|
Promega |
HMS174 (DE3) |
RecA-mutated K-12 background E. coli strain that carries a chromosomal copy of the T7 RNA polymerase gene under the control of the lacUV5 promoter; used as the host strain for the pET expression system |
Novagen |
Plasmids |
|
|
pHPΩ45 |
Contains a 2.0-kb Sm/Spr gene cassette |
33 |
pET30a |
pBR322-derived expression vector with T7 lac promoter and up- and down-stream His tags; Kmr
|
Novagen |
pDMS197 |
A suicide vector; R6K ori sacB Tcr
|
10 |
pALTER-1 |
Vector for the site mutagenesis; Tcr Aps
|
Promega |
pBReno |
The coding region of A. hydrophila enolase gene was cloned in pBR322 at the ScaI/PstI sites and under the control of the Ap promoter of the vector; Tcr
|
This study |
pETUD |
pET30a vector containing up- and downstream DNA flanking sequences to the enolase gene; Kmr
|
This study |
pETUDSm/Sp |
The Sm/Sp cassette was ligated with the up- and downstream DNA flanking sequences to the enolase gene in pET30a vector; Sm/Spr Kmr
|
This study |
pBUDSm/Sp |
The Sm/Sp cassette was ligated with the up- and downstream DNA flanking sequences to the enolase gene in pBluescript vector; Sm/Spr Apr
|
This study |
pDMSUDSm/Sp |
Suicide vector pDMS197 containing the Sm/Sp cassette ligated with up- and downstream DNA flanking sequences to the enolase gene; used to generate the enolase gene deletion mutant of A. hydrophila; Kmr Tcr
|
This study |
pALTER-1/eno |
The native enolase gene of A. hydrophila was cloned into pALTER-1 vector at the BamHI/HindIII sites for the site mutagenesis; Tcr Aps
|
This study |
pALTER/eno343K/Q |
pALTER-1 vector containing the mutated A. hydrophila enolase gene eno(K343Q); Tcs Apr
|
This study |
pALTER/eno394K/M |
pALTER-1 vector containing the mutated A. hydrophila enolase gene eno(K394M); Tcs Apr
|
This study |
pALTER/eno420K/L |
pALTER-1 vector containing the mutated A. hydrophila enolase gene eno(K420L); Tcs Apr
|
This study |
pALTER/eno427K/N |
pALTER-1 vector containing the mutated A. hydrophila enolase gene eno(K427N); Tcs Apr
|
This study |
pALTER/eno430K/R |
pALTER-1 vector containing the mutated A. hydrophila enolase gene eno(K430R); Tcs Apr
|
This study |
pET30aeno |
The native enolase gene of A. hydrophila was cloned in pET30a and fused with the downstream His tags of the vector for hyperexpression; Kmr
|
This study |
pET30aeno343K/Q |
The mutated enolase gene of A. hydrophila eno(K343Q) was cloned in pET30a and fused with the downstream His tags of the vector for hyperexpression; Kmr
|
This study |
pET30aeno394K/M |
The mutated enolase gene of A. hydrophila eno(K394M) was cloned in pET30a and fused with the downstream His tags of the vector for hyperexpression; Kmr
|
This study |
pET30aeno420K/L |
The mutated enolase gene of A. hydrophila eno(K420L) was cloned in pET30a and fused with the downstream His tags of the vector for hyperexpression; Kmr
|
This study |
pET30aeno427K/N |
The mutated enolase gene of A. hydrophila eno(K427N) was cloned in pET30a and fused with the downstream His tags of the vector for hyperexpression; Kmr
|
This study |
pET30aeno430K/R |
The mutated enolase gene of A. hydrophila eno(K430R) was cloned in pET30a and fused with the downstream His tags of the vector for hyperexpression; Kmr
|
This study |