04/12/14
IDP4U
Submitted to: Mrs. Bellissimo
Submitted by: Barento Abdushekur
Jalani Bennett
Daniel Casalinovo
Rebecca Fadoju
Simon Wong
Background Information:
Genetic transformation is taking genes from one organism and putting them into another. Genetic transformation is also when a cell takes up (takes inside) and expresses a new piece of genetic material. Genetic transformation literally means change caused by genes and involves the insertion of one or more genes into an organism in order to change the organism’s traits. A gene is a piece of DNA that gives the instructions for making a protein. This protein gives an organism a certain trait. A gene is inserted into an organism in order to change the organism’s trait. This new genetic information often provides the organism with a new trait which is identifiable after transformation.
Genetic transformation is used in many areas of biotechnology. In agriculture, genes coding for traits such as frost, or drought resistance can be genetically transformed into plants, therefore allowing the plants to take on those traits. In bioremediation, bacteria can be genetically transformed with genes enabling them to digest oil spills. This was used to great effect during the Gulf Oil Spill. In medicine, genes are treated with gene therapy, genetically transforming a sick person’s cells with healthy copies of the defective gene that causes their disease. Genes can be taken out of many sources, such as human, animal, or plant DNA, and placed inside bacteria. For example, a healthy human gene for the hormone insulin can be put into bacteria. Under the right conditions, these bacteria can make authentic human insulin. By doing this, scientists have created a viable treatment for diabetes. Although this cannot cure diabetes, it is a highly effective management of diabetes. In this lab bacteria will be transformed to contain a gene that codes for Green Fluorescent Protein (GFP). The real-life source of this gene is a jellyfish called Aequorea Victoria. GFP causes certain areas of the jellyfish to glow in the dark. The gene for GFP was originally isolated from the jellyfish, Aequorea victoria. The wild-type jellyfish gene has been modified by Maxygen Inc., a biotechnology company in Santa Clara, California. After transforming the bacteria, they will express their new jellyfish gene and produce the fluorescent protein. It causes them to glow a brilliant green color under ultraviolet light. Plasmid DNA usually comprises genes for one or more traits that may be useful to bacterial continued existence. In nature, bacteria can transfer plasmids in and out of a cell, therefore allowing them to share these beneficial genes. The plasmid is a circular, autonomously replicating piece of DNA.
Purpose:
To transform E. coli bacteria by adding plasmids that allow the bacteria to glow green under UV light in the presence of arabinose sugar and grow in the presence of the antibiotic, ampicillin.
Hypothesis:
The bacteria with +pGLO plasmids are resistant to the antibiotic ampicillin and have the gene for GFP, therefore they will survive and grow on the transformation plates that have LB/amp.
+pGLO bacteria on a plate with LB/amp/ara will grow and glow green under UV light because of the inclusion of arabinose.
Materials Student Workstation
E. coli HB101 K–12, lyophilized
4 poured agar plates (1 LB, 2 LB/amp, 1 LB/amp/ara)
Transformation solution
LB nutrient broth
7 Incoculation loops
5 Pipets
Foam microtube holder/float
Container full of crushed ice (foam cup)
Marking pen
Copy of quick guide
Materials at Instructor’s Workstation
1 vial of Rehydrated pGLO plasmid
42 water bath and thermometer
37 incubator
Lesson 1 Focus Questions
1. To genetically transform an entire organism, you must insert the new gene(s) into every cell in the organism. Which organism is better suited for total genetic transformation–one composed of many cells, or one composed of a
