Assay for an
Antibiotic
Resistance Enzyme
Carolina Kit
Focus
Color Assays, pH
Background Info.
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What is an Enzyme
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There are many types of enzymes and each is assigned a
specific task. Enzymes are basically proteins which bring about
a chemical reactions to maintain balance in body functions.
In this lab the enzymes being used are antibiotic resistance
enzymes produced by ampicillin-resistant bacteria.
Background Info.
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Antibiotic resistance in bacteria
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Antibiotic resistance is a type of drug resistance where
a microorganism is able to survive exposure to an antibiotic.
Background Info.
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Genes that confer resistance can easily be
transferred between bacteria, through
conjugation, transduction, or
transformation.
Video
Background Info.
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Enzyme assays
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Are Laboratory methods or tests for measuring
enzymatic activity
Colorimetric assays
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Utilizing color/color change in order to
see enzymatic activity
Timeline/Overview
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Monday—Lecture
Tuesday—pre-lab quiz, Part 1
Thuesday--Part 2 (volunteer to serve as
demonstrator—4 hours lab service)
Monday--handouts due, post quiz
Prep for Lab.
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2 days prior
--label tubes
control (8--15mL tubes)
pAMP (8--15 mL tubes)
penicillin-G (8--15 mL tubes)
--streak fresh plate of MM294 and MM294/pAMP (24 hours at 37 incubator)
1 day prior
--prepare cultures (1/group)—see teacher guide (24 hours at 37 incubator)
--penicillin-G solution (store at -20 until use)
--aliquot (8 groups)
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MM294 5ml overnight culture, label control
MM294/pAMP 5ml overnight culture, label pAMP
5ml of 4mg/mL penicillin-G
On lab day 1
--test the reagents with phenol red
Lab day 2
--demonstrator of spectrometer
Overview and tips
(Tuesday)
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Use correct plate streaking technique
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Make sure incubation is set
to 37 degrees C.
Overview and tips
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When inoculating use Aseptic technique
Don’t forget to
check the temperature
Should be set to
37 degrees C
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