Host Mutation Descriptions:
ara Inability to utilize arabinose.
deoR Regulatory gene that allows for constitutive synthesis for genes involved in
deoxyribose synthesis. Allows for the uptake of large plasmids.
endA DNA specific endonuclease I. Mutation shown to improve yield and quality of DNA
from plasmid minipreps.
F' F' episome, male E. coli host. Necessary for M13 infection.
galK Inability to utilize galactose.
galT Inability to utilize galactose.
gyrA Mutation in DNA gyrase. Confers resistance to nalidixic acid.
hfl High frequency of lysogeny. Mutation increases lambda lysogeny by inactivating specific
protease.
lacI Repressor protein of lac operon. LacIq is a mutant lacI that overproduces the
repressor protein.
lacY Lactose utilization; galactosidase permease (M protein).
lacZ b-D-galactosidase; lactose utilization. Cells with lacZ mutations produce white
colonies in the presence of X-gal; wild type produce blue colonies.
lacZdM15 A specific N-terminal deletion which permits the a-complementation segment present
on a phagemid or plasmid vector to make functional lacZ protein.
Dlon Deletion of the lon protease. Reduces degradation of b-galactosidase fusion proteins
to enhance antibody screening of l libraries.
malA Inability to utilize maltose.
proAB Mutants require proline for growth in minimal media.
recA Gene central to general recombination and DNA repair. Mutation eliminates general
recombination and renders bacteria sensitive to UV light.
rec BCD Exonuclease V. Mutation in recB or recC reduces general recombination to a hundredth
of its normal level and affects DNA repair.
relA Relaxed phenotype; permits RNA synthesis in the absence of protein synthesis.
rspL 30S ribosomal sub-unit protein S12. Mutation makes cells resistant to streptomycin.
Also written strA.
recJ Exonuclease involved in alternate recombination pathways of E. coli.
strA See rspL.
sbcBC Exonuclease I. Permits general recombination in recBC mutants.
supE Supressor of amber (UAG) mutations. Some phage require a mutation in this gene in order
to grow.
supF Supressor of amber (UAG) mutations. Some phage require a mutation in this gene in order
to grow.
thi-1 Mutants require vitamin B1(thiamine) for growth on minimal media.
traD36 mutation inactivates conjugal transfer of F' episome.
umuC Component of SOS repair pathway.
uvrC Component of UV excision pathway.
xylA Inability to utilize xylose.
dam DNA adenine methylase/ Mutation blocks methylation of Adenine residues in the recognition
sequence 5'-G*ATC-3' (*=methylated)
dcm DNA cytosine methylase/Mutation blocks methylation of cytosine residues in the recognition
sequences 5'-C*CAGG-3' or 5'-C*CTGG-3' (*=methylated)
hsdM E. coli methylase/ Mutation blocks sequence specific methylation AN6*ACNNNNNNGTGC or
GCN6*ACNNNNNNGTT (*=methylated). DNA isloated from a HsdM- strain will be restricted by a HsdR+
host.
hsd R17 Restriction negative and modification positive.
(rK-, mK+) Allows cloning of DNA without cleavage by endogenous restriction endonucleases. DNA
prepared from hosts with this marker can efficiently transform rK+ E. coli hosts.
hsdS20 Restriction negative and modification negative.
(rB-, mB-) Allows cloning of DNA without cleavage by endogenous restriction endonucleases . DNA
prepared from hosts with this marker is unmethylated by the hsdS20 modificationsystem.
mcrA E. coli restriction system/ Mutation prevents McrA restriction of methylated DNA of
sequence 5'-C*CGG (*=methylated).
mcrCB E. coli restriction system/ Mutation prevents McrCB restriction of methylated DNA of
sequence 5'-G5*C, 5'-G5h*C, or 5'-GN4*C (*=methylated).
mrr E. coli restriction system/ Mutation prevents Mrr restriction of methylated DNA of sequence
5'-G*AC or 5'-C*AG (*=methylated). Mutation also prevents McrF restriction of methylated cytosine
sequences.
Other Descriptions:
cmr Chloramphenicol resistance
kanr Kanamycin resistance
tetr Tetracycline resistance
strr Streptomycin resistance
D Indicates a deletion of genes following it.
Tn10 A transposon that normally codes for tetr
Tn5 A transposon that normally codes for kanr
spi- Refers to red-gam- mutant derivatives of lambda defined by their ability to form
plaques on E. coli P2 lysogens.
Commonly used bacterial strains
C600 - F-, e14, mcrA, thr-1 supE44, thi-1, leuB6, lacY1, tonA21, l-
-for plating lambda (gt10) libraries, grows well in L broth, 2x TY, plate on NZYDT+Mg.
-Huynh, Young, and Davis (1985) DNA Cloning, Vol. 1, 56-110.
DH1 - F-, recA1, endA1, gyrA96, thi-1, hsdR17 (rk-, mk+), supE44, relA1, l-
-for plasmid transformation, grows well on L broth and plates.
-Hanahan (1983) J. Mol. Biol. 166, 557-580.
XL1Blue-MRF' - D(mcrA)182, D(mcrCB-hsdSMR-mrr)172,endA1, supE44, thi-1, recA, gyrA96, relA1,
lac, l-, [F'proAB, lac IqZDM15, Tn10 (tetr)] -For plating or glycerol stocks, grow in LB
with 20 mg/ml of tetracycline. For transfection, grow in tryptone broth containing 10 mM
MgSO4 and 0.2% maltose. (No antibiotic--Mg++ interferes with tetracycline action.) For picking
plaques, grow glycerol stock in LB to an O.D. of 0.5 at 600 nm (2.5 hours?). When at 0.5, add
MgSO4 to a final concentration of 10 mM.
SURE Cells - Stratagene - e14(mcrA), D(mcrCB- hsdSMR-mrr)171, sbcC, recB, recJ, umuC::Tn5
(kanr), uvrC, supE44, lac, gyrA96, relA1, thi-1, end A1[F'proAB, lacIqDM15, Tn10(tetr)].
An uncharacterized mutation enhances the a - complementation to give a more intense blue color
on plates containing X-gal and IPTG.
GM272 - F-, hsdR544 (rk-, mk-), supE44, supF58, lacY1 or 苐acIZY6, galK2, galT22, metB1m, trpR55,
l-
-for plasmid transformation, grows well in 2x TY, TYE, L broth and plates.
-Hanahan (1983) J. Mol. Biol. 166, 557-580.
HB101 - F-, hsdS20 (rb-, mb-), supE44, ara14, galK2, lacY1, proA2, rpsL20 (strR), xyl-5, mtl-1,
l-, recA13, mcrA(+), mcrB(-)
-for plasmid transformation, grows well in 2x TY, TYE, L broth and plates.
-Raleigh and Wilson (1986) Proc. Natl. Acad. Sci. USA 83, 9070-9074.
JM101 - supE, thi, ?lac-proAB), [F', traD36, proAB, lacIqZ芃15], restriction: (rk+, mk+), mcrA+
-for M13 transformation, grow on minimal medium to maintain F episome, grows well in 2x TY,
plate on TY or lambda agar.
-Yanisch-Perron et al. (1985) Gene 33, 103-119.
XL-1 blue recA1, endA1, gyrA96, thi, hsdR17 (rk+, mk+), supE44, relA1, l-, lac, [F', proAB,
lacIqZ芃15, Tn10 (tetR)]
-for M13 and plasmid transformation, grow in 2x TY + 10 礸/ml Tet, plate on TY agar + 10 礸/ml
Tet (Tet maintains F episome).
-Bullock, et al. (1987) BioTechniques 5, 376-379.
GM2929 - from B. Bachman, Yale E.coli Genetic Stock Center (CSGC#7080); M.Marinus strain; sex F-;
(ara-14, leuB6, fhuA13, lacY1, tsx-78, supE44, [glnV44], galK2, galT22, l-, mcrA, dcm-6, hisG4,[Oc],
rfbD1, rpsL136, dam-13::Tn9, xyl-5, mtl-1, recF143, thi-1, mcrB, hsdR2.)
MC1000 - (araD139, D[ara-leu]7679, galU, galK, D[lac]174, rpsL, thi-1). obtained from the McCarthy
lab at the University of Oklahoma.
ED8767 (F-,e14-[mcrA],supE44,supF58,hsdS3[rB-mB-], recA56, galK2,galT22,metB1, lac-3 or lac3Y1 -
obtained from Nora Heisterkamp and used as the host for abl and bcr cosmids.