Strains
|
Genotype
| |
S. meliloti
| | |
GMI211
|
SmR Nod+ Fix+
|
[20]
|
GMI401
|
GMI211 asnO::Tn5 SmR NmR
|
This work
|
GMI5704
|
GMI211 fixJ2.3::Tn5 SmR NmR BleoR
|
[18]
|
E. coli
| | |
ER
|
asnA31 asnB32 thi
|
[15]
|
MM294
|
Pro-82 thi-1 hsdR17 supE44 endA1
|
[21]
|
DH5 α
|
endA1 hsdR17 (rk-mk+) supE44 thi-1 recA1 gyrA relA1Δ (lacIZYA-
|
[29]
|
|
argF)U169 deoR (φ 80dlacΔ (lacZ)M15)
| |
Plasmids
| | |
pMF457
|
pGD926 (IncP broad host range vector) derivative carrying a fixK-
| |
|
lacZ fusion with a mutation in the fixT promoter. TcR
| |
pCHK57
|
pGMI41211 (Inc-P1 broad host range vector) derivative carrying a
|
[31]
|
|
nifA-lacZ fusion. TcR
| |
pMF11
|
Same as pMF10 but with fixT in reverse orientation. GmR
| |
pBH1
|
pBBR1-MCS3 derivative, carrying an HindIII fragment containing
|
This work
|
|
the asnO gene and the SMb20482 orf.
| |
pBasn2
|
pBBR1-MCS3 derivative, expressing asnO under the control of the
|
This work
|
|
pLac promoter
| |
pUC23
|
pUC18 digested with EcoRI, carrying the S. meliloti EcoRI
|
This work
|
|
fragment containing the asn O region with the Tn5 insert. ApR,
| |
|
KmR.
| |
BAC37
|
pBeloBAC11 containing a 100 kb S. meliloti genome fragment
|
[26]
|
|
carrying the asn-like region of pSymB.
| |
pRK602
|
PRK600::Tn5, Cmr, Nm-Kmr
|
[21]
|