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Historia de los trenes de E.

coli
Las E. coli son bacterias gramnegativas con forma de bastón que recibieron su nombre del Dr.
Theodor Escherich, el científico que las describió por primera vez en 1885. Las E. coli se
encuentran principalmente en el tracto intestinal de los animales. Hay muchas cepas diferentes
de E. coli de origen natural , algunas de las cuales son mortales para los humanos. La mayoría
de las cepas de laboratorio comerciales comunes de E. coliutilizados hoy en día descienden de
dos aislados individuales, la cepa K-12 y la cepa B. K-12 se aisló de un paciente en 1920 y
finalmente condujo a las cepas de laboratorio comunes MG1655 y sus derivados DH5alpha y
DH10b (también conocidos como TOP10). La historia de la cepa B es un poco más complicada
debido a que los investigadores comparten y cambian el nombre de las muestras a lo largo de
la historia. Probablemente se aisló en 1918, pero se denominó por primera vez "cepa B" en
1942. La cepa BL21 y sus derivados son los ejemplos más comunes de la cepa B de E. coli .

Trenes de E. coli comunes utilizados en el


laboratorio
La mayoría de las cepas comerciales que se encuentran en la actualidad se comercializan con
un propósito específico: crecimiento rápido, clonación de alto
rendimiento, clonación de rutina, clonación de ADN inestable,
preparación de ADN no metilado y más. Muchas mutaciones
que hacen posibles estas características están presentes en la
mayoría de las cepas comerciales, especialmente las
mutaciones que hacen mejoras importantes, como las que
aumentan el rendimiento del plásmido y / o la calidad del
ADN. La Tabla 1 a continuación describe algunos de los
cambios genéticos más comunes que se encuentran en
las cepas de E. coli .

Tabla 1: Mutaciones genéticas comunes encontradas en cepas


de E. coli

Gene(s) Description Functional Consequence


Preparing unmethylated DNA, important when trying
DNA adenine methylase mutation
dam to cut with certain restriction enzymes (ex: ClaI or
(GATC)
XbaI)
Preparing unmethylated DNA, important when trying
DNA cytosine methylase mutation
dcm to cut with certain restriction enzymes that
(CCWGG)
are methylation sensitive.
dnaJ Mutation in a chaperonin gene Increases the stability of certain expressed proteins
endA, Endonuclease I (nonspecific cleavage
Improves plasmid yield
endA1 of dsDNA ) mutation
A low copy-number plasmid, encodes proteins
Host does (F') or does not (F-) contain
F needed for bacterial conjugation. Genes listed on F´
the fertility plasmid.
are wild-type unless indicated otherwise
fhuA
ferric hydroxamate uptake, iron uptake
(formerly T1/T5 Phage resistance
receptor mutation.
tonA)
Mutation in galactose metabolism
gal Cells cannot grow on galactose only
pathway
gyrA,
DNA gyrase mutation Confers resistance to nalidixic acid
gyrA96
Unmethylated DNA not degraded, cell still can
hsdRMS hsdR(rk-, mk+)
methylate DNA
Unmethylated DNA not degraded, cell cannot
hsdS(rk-,mk-)
methylate DNA
lac Lac operon mutations Blue/white screening of clones
lac repressor overproduced, expression from pLac
lacIq
repressed more
LacZ β-galactosidase activity abolished
Lactose permease inactivated, lactose cannot be
lacY
taken up by cell
mcrA, Inactivation of pathway that cleaves
Allows for uptake of foreign (methylated) DNA
mcrBC methylated cytosine DNA
mrr, Inactivation of pathway that cleaves
Allows for uptake of foreign (methylated) DNA
Δ(mcrC-mrr) methylated adenine or cytosine DNA
Mutation in a DNA-dependent ATPase
recA, recA1, Reduces plasmid recombination, increases plasmid
that is essential for recombination and
recA13 stability
general DNA repair
recBCD Exonuclease V activity abolished Increased plasmid stability, reduced recombination
Relaxed phenotype, mutation Allows RNA synthesis in absence of protein
relA or relA1
eliminating stringent factor synthesis
Ptrc-ccdA Propagation of ccdB-containing plasmids
Hte High transformation efficiency
constitutive expression of genes for
deoR Allows uptake of large plasmids
deoxyribose synthesis
hee High electroporation efficiency
supE44 Suppression of the amber (UAG) stop codon by
(glnV44) glutamine insertion
Suppression of the amber (UAG) stop codon by
supF (tyrT)
tyrosine insertion
λ-thi-1 or
Mutation in thiamine metabolism Requires exogenous thiamine for growth
thi1
disruption of arabinose metabolism
ara Inability to utilize arabinose as a carbon source
pathway
β-isopropyl malate dehydrogenase
leuB Requires exogenous leucine source for growth
inactivated
mutation in proline biosynthesis
proAB Requires exogenous proline source for growth
pathway
Mutation in subunit S12 of 30S
rpsL Confers resistance to streptomycin
ribosome
Tn10 Confers resistance to tetracycline
Además, la Tabla 2 proporciona una referencia rápida para algunas de las cepas populares,
sus genotipos y su uso principal en el laboratorio. Todas estas cepas están basadas en E.
coli K-12 y se consideran BSL-1.

Tabla 2: Cepas de laboratorio de E. coli

Natural
Strain Primary Use Genotype
resistance
TOP10 derivative. For F-mcrA Δ(mrr-hsdRMS-mcrBC)
ccdB
propagating plasmids Φ80lacZΔM15 ΔlacX74 recA1 araΔ139
Survival 2
expressing the ccdB gene Δ(ara-leu)7697 galU galK rpsL
T1R
(important in Gateway cloning). (StrR) endA1 nupG fhuA::IS2
HB101 derivative. For F- gyrA462 endA1 glnV44 Δ(sr1-recA) mcrB
propagating plasmids mrr hsdS20(rB-, mB-) ara14 galK2 lacY1
DB3.1 Streptomycin
expressing the ccdB gene proA2 rpsL20(Smr) xyl5 Δleu mtl1
(important in Gateway cloning).
F- endA1 recA1 galE15 galK16 nupG rpsL
MC1061 derivative. General
ΔlacX74 Φ80lacZΔM15 araD139
DH10B Streptomycin cloning and storage, blue/white
Δ(ara,leu)7697 mcrA Δ(mrr-hsdRMS-
screening, leucine auxotroph.
mcrBC) λ-
General cloning and storage of F- endA1 glnV44 thi-1 recA1 relA1 gyrA96
DH5alpha common plasmids, blue/white deoR nupG Φ80dlacZΔM15 Δ(lacZYA-
screening. argF)U169, hsdR17(rK- mK+), λ–
Hybrid of E. coli K12 and E. coli
B (mostly K12, though), F- mcrB mrr hsdS20(rB- mB-) recA13 leuB6
HB101 Streptomycin common lab strain for cloning ara-14 proA2 lacY1 galK2 xyl-5 mtl-1
and storage of pBR322 rpsL20(SmR) glnV44 λ-
plasmids.
General cloning and plasmid
maintenance, blue/white endA1 glnV44 thi-1 relA1 gyrA96 recA1
JM109 screening, partly restriction- mcrB+ Δ(lac-proAB) e14- [F' traD36
deficient; good strain for cloning proAB+ lacIq lacZΔM15] hsdR17(rK-mK+)
repetitive DNA.
For storing plasmids that
should not be Dam or Dcm rpsL thr leu thi lacY galK galT ara tonA tsx
methylated, allows for dam dcm glnV44 Δ(lac-proAB) e14- [F'
JM110 Streptomycin
methylation sensitive restriction traD36 proAB+ lacIq lacZΔM15] hsdR17(rK-
enzymes to cut the plasmid mK+)
after preparation.
Parent of DH10B/TOP10 and F- Δ(ara-leu)7697 [araD139]B/r Δ(codB-lacI)3
MC1061 Streptomycin derived strains, common lab galK16 galE15 λ- e14- mcrA0 relA1
cloning and storage strain. rpsL150(strR) spoT1 mcrB1 hsdR2(r-m+)
Wild type K-12 strain; first
MG1655 published sequence of an E. F- λ- ilvG- rfb-50 rph-1
coli K-12 strain.
For cloning into and storage of
F' proA+B+ lacIq ∆(lacZ)M15
lentiviral and retroviral vectors
NEB zzf::Tn10 (TetR) ∆(ara-leu) 7697 araD139
or cloning or repeated
Stable fhuA ∆lacX74 galK16 galE15
sequences with the potential to
e14- Φ80dlacZ∆M15 recA1 relA1 endA1
recombine.
nupG rpsL (StrR) rph spoT1 ∆(mrr-hsdRMS-
mcrBC)
For cloning and maintenance of
a plasmids with R6Kγ origin;
contains a mutant allele of the F- ∆lac169 rpoS(Am) robA1 creC510
Pir1
pir gene that maintains the hsdR514 endA recA1 uidA(∆MluI)::pir-116
plasmids at ~250 copies per
cell.
JM109-derived. For cloning into
and storage of lentiviral and F- endA1 glnV44 thi-1 recA1 gyrA96 relA1
Stbl2 retroviral vectors or cloning or Δ(lac-proAB) mcrA Δ(mcrBC-hsdRMS-mrr)
repeated sequences with the λ-
potential to recombine.
Derived from HB101. For
cloning into and storage of
lentiviral and retroviral vectors
or cloning or repeated F- glnV44 recA13 mcrB mrr hsdS20(rB-,
Stbl3 Streptomycin sequences with the potential to mB-) ara-14 galK2 lacY1 proA2 rpsL20 xyl-5
recombine. This strain is leu mtl-1
endA+, so column-based DNA
preps require an additional
wash step.
Electrocompetent cells for
F' mcrA Δ(mcrBC-hsdRMS-
cloning and storage of unstable
mrr) recA1 endA1 gyrA96 gal-thi-1 supE44 λ-
Stbl4 DNA and lentiviral/retroviral
relA1 Δ(lac-
vectors. Used by Addgene
proAB)/F' proAB+lacIqZΔM15 Tn10 (TetR)
for pooled library amplification.
F- mcrA Δ(mrr-hsdRMS-mcrBC)
MC1061 derivative. General
φ80lacZΔM15 ΔlacX74 nupG recA1
TOP10 Streptomycin cloning and storage, blue/white
araD139 Δ(ara-leu)7697 galE15 galK16
screening.
rpsL(StrR) endA1 λ-
Blue/white screening and endA1 gyrA96(nalR) thi-1 recA1 relA1 lac
XL1 Blue Tetracycline routine cloning, nalidixic acid glnV44 F'[ ::Tn10 proAB+ lacIq Δ(lacZ)M15]
resistant. hsdR17(rK- mK+)
High competency cloning and endA1 glnV44 recA1 thi-1 gyrA96 relA1 lac
XL10 Tetracycline and propagation of large plasmids, Hte Δ(mcrA)183 Δ(mcrCB-hsdSMR-mrr)173
Gold Chloramphenicol ligated DNA, and libraries, tetR F'[proAB lacIqZΔM15 Tn10(TetR Amy
nalidixic acid resistant. CmR)]

Nota: Las mutaciones inactivadoras en genes específicos se indican con un signo menos (-)
como suele ser estándar; tener el gen en la lista indica que no es funcional. Si se elimina un
gen, normalmente se indica con un delta griego (Δ).

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