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Transformation of E. coli Using Green Fluorescent Protein


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Transformation of E. coli Using Green Fluorescent Protein

In this activity related to plant biotechnology, learners transform a strain of E. coli using green fluorescent protein from a bioluminescent jellies. This lab will help learners understand recombinant DNA techniques, including the transformation procedure using the heat shock method, the uses of marker or reporter genes in molecular biology experiments and how to screen for a gene of interest, how DNA can be transferred to another organism and therefore change the observable characteristics of that organism, and how to calculate transformation efficiency. Learners will also become familiar with sterile technique and decontamination procedures to safely handle bacteria. Although the E. coli strain used in the experiment is non-pathogenic, it's important to emphasize proper safety precautions and adult supervision is highly recommended. Modifications for younger learners are included in a related PDF (see related resources). This resource also contains background information, safety tips, and questions with answers for learners.

Quick Guide


Preparation Time:
1 to 7 days

Learning Time:
1 to 7 days

Estimated Materials Cost:
Over $20 per group of students

Age Range:
Ages 11 - adult

Resource Types:
Activity, Demonstration, Experiment/Lab Activity, Lesson/Lesson Plan

Language:
English

Materials List (per group of students)


  • Disposable gloves
  • Safety goggles
  • Waterproof marker
  • Microtube rack
  • 6 disposable pipettes
  • 6 disposable inoculating loops
  • Bacterial waste container
  • Foam cup with crushed ice
  • 2 plates with LB Medium
  • 2 plates with LB medium + ampicillin
  • Microtube filled with 50 mM CaCl2
  • Microtube filled with Luria-Bertani broth
  • Squirt bottles containing 10% bleach
  • 20-µl micropipette and tips for instructor use only
  • Water bath (with floating tube racks)
  • Plates with E. coli cells streaked out and grown overnight
  • pGREEN plasmid (0.005 µg/µl)
  • Crushed ice
  • Distilled water
  • 37°C incubator
  • Parafilm
  • UV light
  • Container with 10% bleach for sterilization of all items that come into contact with the bacteria
  • You will need a large container with 10% bleach solution to contain all used disposable pipets and loops and to sterilize the petri dishes

Subjects


  • Engineering and Technology
    • Engineering
      • Bioengineering/Biomedical Engineering
    • Technology
      • Agriculture and Biotechnology
  • Life Sciences
    • Cells
      • Cell Structure and Function
      • Cell Division
      • Chemistry of Life
    • Diversity of Life
      • Animals
      • Viruses and Bacteria
    • Heredity and Genetics
      • DNA Structure and Function
      • Genetic Engineering
  • Mathematics
    • Data Analysis and Probability
      • Data Analysis
      • Data Collection
      • Data Representation
    • Number and Operations
  • Physical Sciences
    • Chemistry
      • Chemistry of Life
    • Light and Optics
  • The Nature of Science
    • The Scientific Process
      • Conducting Investigations
      • Gathering Data
      • Formulating Explanations
      • Communicating Results

Informal Categories


  • Animals

Audience


To use this activity, learners need to:

  • see
  • see color
  • read
  • touch

Learning styles supported:

  • Involves teamwork and communication skills
  • Involves hands-on or lab activities

Other


Components that are part of this resource:

This resource is part of:

Access Rights:

  • Free access

By:

  • Stephens, Janice; Leach, Jan

Rights:

  • All Rights Reserved, The American Phytopathological Society, ©2011

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