Assay for an Antibiotic Resistance Enzyme
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Transcript Assay for an Antibiotic Resistance Enzyme
Assay for an
Antibiotic
Resistance Enzyme
Carolina Kit
Focus
Color Assays, pH
Background Info.
What is an Enzyme
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.
Antibiotic resistance in bacteria
Antibiotic resistance is a type of drug resistance where
a microorganism is able to survive exposure to an antibiotic.
Background Info.
Genes that confer resistance can easily be
transferred between bacteria, through
conjugation, transduction, or
transformation.
Video
Background Info.
Enzyme assays
Are Laboratory methods or tests for measuring
enzymatic activity
Colorimetric assays
Utilizing color/color change in order to
see enzymatic activity
Timeline/Overview
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.
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)
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)
Use correct plate streaking technique
Make sure incubation is set
to 37 degrees C.
Overview and tips
When inoculating use Aseptic technique
Don’t forget to
check the temperature
Should be set to
37 degrees C