Gene Cloning and Transgenic Organism Flashcards
(24 cards)
What is gene cloning
Process of isolating and copying a gene of interest to facilitate its expression within a host organism
What are the 3 key stages of gene cloning
Isolation of gene and vector
Insertion of gene into vector
Integration of recombinant vector into host cell
What is a vector in gene cloning
A molecule like a plasmid used to carry the gene of interest into host cell
What role do restriction enzymes play in gene cloning
They cut DNA at specific sequences, producing sticky or blunt ends for ligation
What is the function of DNA ligase
Joins gene of interest to vector by forming phosphodiester bonds
What is transformation in gene cloning
Process of introducing the recombinant vector into a host like bacteria
How can successful transformation be confirmed
By using antibiotic selection markers, fluorescent protein, enzyme
What is the purpose of using reverse transcriptase in gene cloning
Synthesise complementary (cDNA) from mRNA without introns
What is the significance of sticky ends in gene cloning
Allow for complimentary base pairing, facilitating the ligation of DNA fragments
What is the role of a bacterial plasmid in gene cloning
Serves as a vector to carry gene of interest to host cell
What is transgenic organism
One that has received and expressed recombinant DNA
What is cloning vector
DNA molecule used to introduce foreign DNA
What are the steps involved in making transgenic organism
Isolate DNA
Cut DNA from chromosome
Make millions of copies of it
Put DNA into vector
Put DNA into host and turn on gene expression
How is gene cloning used in gene therapy
Allows creation of therapeutic gene for genetic disorders
Why is successful gene therapy difficult
Targeting correct cells
Ensuring long term expression
Avoiding immune reactions
Potential effects like cancer
What is the nuclear transfer technique
Method where nucleus of somatic cell is transferred into an enucleated egg cell to create clone
How does nuclear transfer differ from gene cloning
NT produces an entire cloned organism
GC focuses on copying specific genes to express them in host cells
What is a promoter sequence in a transgene
Initiates transcription of the transgene in target cell
What is a poly-A in a transgene
DNA sequence that ensures proper mRNA processing and stability
What is in vivo cloning
Inside living organism
What are restriction enzymes (endonucleases)
Enzyme that cut DNA at specific palindromic recognition sites - produce sticky/blunt ends
What’s transfection
Introduction of foreign DNA into eukaryotes
How is heat shock used in transformation
Cells and DNA are chilled, then rapidly heated to promote DNA uptake
What’s electroporation
High voltage pulse that temporarily disrupts cell mem allowing DNA to enter
