[22] Construct Design for Cloning Flashcards
What is the purpose of construct design in cloning?
To create a DNA molecule with specific features for insertion into a host organism.
What elements are typically included in a construct for cloning?
A vector, the gene of interest, a promoter, a terminator, and selectable markers.
What is the role of a vector in a construct for cloning?
It is the DNA molecule that will carry the gene of interest into the host organism.
What is the purpose of the gene of interest in a cloning construct?
It’s the gene that we want to clone and express in the host organism.
What is the function of a promoter in a construct for cloning?
It initiates transcription of the gene.
What is the role of a terminator in a construct for cloning?
It signals the end of transcription.
What is the role of selectable markers in a construct for cloning?
They allow for the identification of cells that have successfully taken up the vector.
What are some examples of selectable markers?
Antibiotic resistance genes and color change genes.
What is the purpose of restriction sites in a cloning construct?
They are specific sequences recognized by restriction enzymes, allowing specific insertion points for the gene of interest.
What is a multiple cloning site (MCS)?
A region in a vector that contains multiple restriction enzyme recognition sites.
MCS - Initials
Multiple Cloning Site
What is the purpose of an origin of replication in a cloning construct?
It allows the vector to replicate within the host organism.
What is a reporter gene?
A gene that is not present in the organism being studied and that is linked to the gene of interest in a cloning experiment.
What is the purpose of a reporter gene in cloning?
It is used to indicate whether the gene of interest has been taken up by the host organism and is being expressed.
What are some examples of reporter genes?
Green Fluorescent Protein (GFP), β-galactosidase (lacZ), and luciferase.
