PROMEGA CORPORATION 2800 WOODS HOLLOW ROAD MADISON, WI 53711-5399 USA TELEPHONE 608-274-4330 www.promega.com TECHNICAL REFERENCE
©2010 ALL RIGHTS RESERVED PART #GE627 Antibiotics S) of and A (Sm) Streptomycin A protein (Kan) Kanamycin by (Cm) A Chloramphenicol A (Amp) Ampicillin Antibiotic Resistance. of Mechanism and Action of Mode Antibiotics: Binds synthesis G418 An (Puro) Puromycin ribosome A (Hygro) Hygromycin by A bacterial (Neo) Neomycin A (Tet) Tetracycline
ribosomes Modeof Action in causing on other to with toxicity High subunit messenger
bactericidal bactericidal bacteriostatic derivative protein bactericidal light-sensitive
eukaryotes ribosomes prokaryotic
interfering binding both aminonucleoside
causes
mito
concentrations
eukaryotic to
to
synthesis
of
prokaryotic
premature the
chondrial protein
synthesis
translocation by eukaryotic
ribosomes. and to
RNA.
misreading
of 80S
specifically
and the with and
agent agent agent ribosomes,
penicillin
preventing
agent bacteriostatic ribosomes. synthesis
prokaryotic by ribosomal
causes
blocks bacterial
ribosomes,
chain inhibitor
and
of that that that binding antibiotic cells
that
neomycin
and
of that
eukaryotic inhibiting
binds binds blocks
protein termination.
misreading
and
because
messenger peptide by interferes
causes cell
subunit
that
to kills
70S
binding
with
agent that which the to to
wall
protein
interferes
ytei. phosphotransferase) synthesis. growing
can the 70S ribosomal
peptidyl
bond
50S blocks reduced it
iooe ribosomes itasain transferase mistranslation.
found
synthesis.
that
with
can are of
30S to result
N. (aminoglycoside RNA. ribosomes
synthesis subunit the the omto. inactivates, formation.
similar prevents
interact
bacterial cells rti The protein
ny Expression only subunit rnfr N-acetyl-transferase. transfer with
in modifies affinity, 30S uui. phosphotransferase) subunit.
The enzyme,
80S of
modifies
The Expression
acetyltransferase The The The
The antibiotic ring Mechanismof Resistance eukaryotic which thereby 4-hydroxyl transport the
ribosome. resistance resistance resistance resistance resistance resistance resistance of
lacks the
producing
β the the and of
-lactamase,
cells antibiotic. group
of of
the that the
biological antibiotic bacterial
the the prevents
gene gene gene gene gene gene gene
antibiotic.
results antibiotic catalyzes phosphotransferase)
on that bacterial bacterial
7
′
the -O-phosphoryl-hygromycin ( ( ( ( ( ( ( gene gene
bla hph tet str kan caf pac
acetylates, membrane activity
and which its in
) ) cyclitol )
)
) )
) into
detoxification specifies specifies
the specifies
interaction
specifies (derived (derived specifies encodes specifies APH APH inhibits
cleaves both phosphorylation the
aiolcsd G418 (aminoglycoside 50µg/ml (aminoglycoside ring
and and cell.
its
in
from from a an
a
uoyi 1–10µg/ml puromycin an an a
(hyosamine),
protein with vivo
eilsi 5–2µ/l 50mg/ml 50–125µg/ml periplasmic
that
binding the thereby rvns in prevents hsh- 50–1,000µg/ml phospho- enzyme
nye 0gm 50mg/ml 30µg/ml enzyme of
Tn5) n) bacterial Tn5).
β and modifies ribosomes. 48 at G418. lca in -lactam
ht 0gm 12.5mg/ml 10µg/ml that
n for in o in to
of in ht 0gm 50mg/ml 30µg/ml that
, for B,
vitro. the
selection;
the
for
mammalian in 12.5µg/ml
0100gm buffered 50–1,000µg/ml Concentration with bacterial 010gm 34mg/ml 20–170µg/ml
initial liquid
in Working mammalian
500µg/ml,
is
a
ethanol
20–200µg/ml
water water water
often range
culture to (pH eeto in selection selection utr; in culture;
in eeto HEPES selection eeto or selection
for plates
o 10mg/ml for
used 50mg/ml
of
25mg/ml
HEPES
prepared in 100mg/ml
water
(pH maintain
HEPES
Solution 7.0) Stock
ethanol
or
water water
media
(pH in
7.0)
or 100mM solution buffer in
buffer a
water tissue
either
7.3)
in highly
pH