Lab 8: Bacterial Transformation

Question 1

Define the term “genetic transformation”.

Answer 1

Genetic tranformation - is the change caused by genes. Involves the insertion of a gene into an organism in order to change the organisms trait.

Question 2

Describe and explain the purpose of the steps in the transformation process.

Answer 2

Steps in Tranformation Process
1. Transfer transformation solution (CaCl2) into a tube where you can mix colonies of bacteria: **This introduces new plasmids which bacteria amplifies making more copies of it. **

2. Heat shock from 0 degrees celsius to 42 degree Celsius: **creates pores where it facillitates the entry of DNA into the bacteria cells. **
3. Now add LB nutrient broth:** LB provides nutrients for bacteria to grow good and fast. **
4. Now are incubated to restore cell wall and cell memebrane.

Question 3

What are plasmids?

Answer 3

Plasmids: small circular DNA fragements found in bacteria cytoplasm.
Used to move genes in and out of bacteria in the lab.

Question 4

What is the purpose of the ampicillin resistance on the plasmid?

Answer 4

The ampicillin resistance on the plasmid purpose is to transfer the resitant gene into a susceptible strain in the bacteria to prevent ampicillin resitance.

But in our experiment is used as a slective marker to see which ones pick up the gene.

Question 5

Analyze and interpret data from a transformation experiment.

Question 6

In our experiment, what does arabinose do? How does it regulate the GFP gene?

Answer 6

The gene for GFP can be switched on in transformed cells by adding the sugar arabinose to the cells’ nutrient medium.

This on is as arabinose promotes gene expression.

GFP is regulated as arabinose promotes the binding of RNA polymerase to the promoter, which causes transcription of the GFP gene into messenger RNA (mRNA), followed by the translation of this mRNA into GFP.

Question 7

What does the transformation efficiency mean? How is it calculated?

Answer 7

Transformation efficiency, gives you an indication of how effective you were in getting DNA molecules into bacterial cells.

Calculated:
TE = total number of succesful transformat/amount of DNA used during the transformation procedure