Vernier Software and Technology
Vernier Software & Technology

pGLO™ Bacterial Transformation

Figure from experiment 6A from Advanced Biology with Vernier

Introduction

Introduction to Transformation

In this lab, you will perform a procedure known as genetic transformation. Genetic transformation literally means “change caused by genes”, and occurs when the cell incorporates and expresses a new piece of genetic material – DNA derived from another organism. Transformation involves the insertion of a gene into an organism in order to alter the recipient organism’s expression. Genetic transformation is used in many areas of biotechnology. In agriculture, genes coding for traits such as frost, pest, or spoilage resistance can be genetically transformed into plants. In bioremediation, bacteria can be genetically transformed with genes enabling them to digest and breakdown pollutants such as oil spills or heavy metals contamination In medicine, disorders caused by defective genes are being treated by gene therapy; that is, by genetically transforming a sick person’s cells with healthy copies of the defective gene.

Your Protein of Interest - The Green Fluorescent Protein, GFP

You will use a procedure to transform bacteria with a gene that codes for Green Fluorescent Protein, GFP. The real-life source of this gene is the bioluminescent jellyfish Aequorea victoria. Following the transformation procedure, the bacteria will express their newly acquired gene and produce GFP, which causes them to glow a brilliant green color under ultraviolet or blue light.

The Host Cell - Escherichia coli (E. coli)

The bacterium, E. coli, is the ideal host for transformation because it is a small, single-celled organism that reproduces quickly, so its transformation will be seen rather quickly. Also, the strain of E. coli being used is nonpathogenic, does not make people or animals sick, and it does not survive outside the laboratory environment. Although it is safe, it requires the use of Standard Microbiological Practices, as directed by your instructor.

Objectives

In this experiment, you will

  • Use a plasmid vector to transform bacteria with genes for Green Fluorescent Protein (GFP) and antibiotic resistance in a controlled experiment.
  • Use the heat shock method of transforming E. coli.
  • Regulate the expression of the GFP gene using arabinose.
  • Describe the biological process involved in transforming bacterial cells.
  • Calculate your transformation efficiency.
  • Learn basic molecular biology techniques.

Sensors and Equipment

This experiment features the following Vernier sensors and equipment.

Additional Requirements

You may also need an interface and software for data collection. What do I need for data collection?

Standards Correlations

See all standards correlations for Advanced Biology with Vernier »

Advanced Biology with Vernier

See other experiments from the lab book.

1ADiffusion through Membranes
1BOsmosis
2AEnzyme Action: Testing Catalase Activity
2BEnzyme Action: Testing Catalase Activity
3Mitosis & Meiosis
4APlant Pigment Chromatography
4BPhotosynthesis
5ACell Respiration (CO2 and O2)
5BCell Respiration (CO2)
5CCell Respiration (O2)
5DCell Respiration (Pressure)
6ApGLO™ Bacterial Transformation
6BAnalysis of Precut Lambda DNA
6BForensic DNA Fingerprinting
7Genetics of Drosophila
8Population Genetics and Evolution
9Transpiration
10ABlood Pressure as a Vital Sign
10BHeart Rate and Physical Fitness
11Animal Behavior
12ADissolved Oxygen in Water
12BPrimary Productivity
13The Visible Spectra of Plant Pigments
14Determination of Chlorophyll in Olive Oil
15Enzyme Analysis using Tyrosinase
16Introduction to Neurotransmitters using AChE
17Macromolecules: Experiments with Protein

Experiment 6A from Advanced Biology with Vernier Lab Book

<i>Advanced Biology with Vernier</i> book cover

Included in the Lab Book

Vernier lab books include word-processing files of the student instructions, essential teacher information, suggested answers, sample data and graphs, and more.

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