Genetic Engineering Selection Techniques

This video tutorial explains how antibiotic resistance genes function as selectable markers in the selection of recombinant colonies. It begins with defining selectable markers and uses the PPR 322 Vector as an example, highlighting its ampicillin and tetracycline resistance regions. The concept of insertional inactivation is introduced, showing how it helps in selecting recombinant colonies. The video describes the transformation experiment, detailing the types of colonies formed and the use of replica plating to identify colonies with the gene of interest. The tutorial concludes with a summary and a call to action for viewers to subscribe.

What is the primary function of a selectable marker in genetic engineering?

• To enhance the growth rate of bacteria

• To identify and select recombinant colonies

• To prevent contamination in cultures

• To increase the size of plasmids

Which two antibiotic resistance regions are present in the PPR 322 Vector?

• Neomycin and Gentamicin

• Ampicillin and Tetracycline

• Streptomycin and Erythromycin

• Kanamycin and Chloramphenicol

What is the role of the BamH1 restriction enzyme in this experiment?

• To replicate the vector

• To cut the vector at a specific site

• To enhance antibiotic resistance

• To transform the bacteria

What is the result of inserting a gene of interest into the tetracycline resistance gene sequence?

• The gene of interest becomes nonfunctional

• The tetracycline resistance gene becomes nonfunctional

• The vector is destroyed

• Both genes become functional

Why is insertional inactivation important in selecting recombinant colonies?

• It inactivates a resistance gene, allowing selection

• It prevents the insertion of the gene of interest

• It makes the vector more stable

• It enhances the growth of all colonies

How many types of colonies are typically observed after a transformation experiment?

• Two

• One

• Three

• Four

Which type of colony can grow in both tetracycline and ampicillin containing media?

• All of the above

• Transformed with recombinant vector

• Transformed with non-recombinant vector

• Non-transformed

What is the purpose of replica plating in the context of this experiment?

• To reduce the number of transformed colonies

• To increase the number of colonies

• To identify colonies with the gene of interest

• To enhance the growth of non-transformed colonies

Which colonies are identified as having the gene of interest using replica plating?

• Colonies that do not grow in any media

• Colonies that grow in tetracycline but not in ampicillin

• Colonies that grow in both media

• Colonies that grow in ampicillin but not in tetracycline

What happens to non-transformed colonies in the presence of antibiotic media?

• They do not grow

• They become resistant

• They transform into recombinant colonies

• They grow normally