21.2 - In Vivo Cloning Flashcards
Inserting a DNA fragment into a vector Using vectors to transfer DNA into host cells Identifying transformed host cells The need for promoter and terminator regions
What is in vivo cloning?
Process of producing large quantities of a target DNA fragment in living cells
What are the key steps in inserting DNA into vectors?
- A vector is cut open at a specific site using a restriction enzyme, creating sticky ends.
- The same restriction enzyme is used to cut the target DNA fragment, creating complementary sticky ends.
- DNA ligase forms phosphodiester bonds between the sugar and phosphate groups on the two strands of DNA, joining the sticky ends of the vector and DNA fragment together.
- The newly formed combined DNA molecule is known as recombinant DNA.
What is transformation?
Introducing vectors with recombinant DNA into host cells
What are plasmid vectors?
Small circular DNA molecules in bacteria
How are host cells treated for plasmid uptake?
Calcium ions
What are bacteriophage vectors?
Viruses that infect bacteria
How do bacteriophage vectors work?
Inject DNA into host
Why is it important to identify transformed host cells?
Only a small percentage uptake DNA
What are marker genes?
Genes indicating which cells took up recombinant DNA
How are marker genes used?
Inserted into vectors
What are examples of marker genes?
Antibiotic resistance
What control elements are needed for protein production?
Promoter regions
What is the function of promoter regions?
Binding area for RNA polymerase
What is the function of terminator regions?
Indicate transcription end
