Bacterial Strains
Please note that the strains DY380, EL250, and EL350 are no longer available.
They were replaced by the new strains SW102, SW105, and SW106, respectively. The
new strains are identical to the old ones, except they have been modified to also
allow for efficient BAC modification using our new galK positive/negative selection
procedure.
DY380 (not available): The original, DH10B-derived strain. These bacteria
contain a defective λ prophage with recombination proteins exo, bet,
and gam being controlled by the temperature-sensitive repressor cI857.
The bacteria are tet resistant (12.5 μg/ml). Genotype: F- mcrA Δ(mrr-hsdRMS-mcrBC)
Φ80dlacZ M15 ΔlacX74 deoR recA1 endA1
araD139 Δ(ara, leu) 7649 galU galK rspL
nupG [ λcI857 (cro-bioA) <> tet]
Reference
EL250 (not available): Derived from DY380. In addition to the recombination
genes found in DY380 EL250 also contains a tightly controlled arabinose-inducible
flpe gene. flpe will mediate recombination between two identical frt
sites. This strain is not tet resistant. Genotype: DY380 [(cro-bioA) <>
araC-PBADflpe]
Reference
EL350 (not available): Derived from DY380. In addition to the recombination
genes found in DY380 EL350 also contains a tightly controlled arabinose-inducible
cre gene. cre will mediate recombination between two identical loxP
sites. This strain is not tet resistant. Genotype: DY380 [(cro-bioA) <>
araC-PBADcre]
Reference
SW102: Derived from DY380 and has all the same features, but designed for
BAC recombineering using galK positive/negative selection. These bacteria
contain a fully functional gal operon, except the galK gene is missing.
The ability to grow on galactose minimal medium can be restored by adding galK
in trans, by inserting a galK expression cassette into a BAC. Next, the galK
cassette can be replaced by any dsDNA carrying any desired mutation, by selection
against galK using 2-deoxy-galactose.
Reference
SW105: Derived from EL250. Like SW102, these cells can be used for galK
positive/negative selection. Like EL250, these cells contain an ara-inducible
Flpe gene.
Reference
SW106: Derived from EL350. Like SW102, these cells can be used for galK
positive/negative selection. Like EL350, these cells contain an ara-inducible
Cre gene.
Reference