recombinant genes Flashcards
(23 cards)
What are recombinant proteins?
Proteins produced by expression of a gene placed into a heterologous system.
Why are recombinant proteins made?
To create exact copies of human proteins for replacement therapy and to produce proteins that do not exist in nature.
What is a heterologous system?
A system where a protein is expressed from a gene not usually found in that host.
What is gene expression?
Placing an in-frame section of open reading frame DNA downstream of an appropriate promoter.
What is the role of microbial bioreactors?
To produce recombinant proteins such as human insulin using prokaryotic systems like E. coli.
What are eukaryotic bioreactors used for?
To produce proteins such as Hepatitis B surface antigen using yeast or insect cells.
What is the significance of mammalian cell-derived bioreactors?
They account for 60% of therapeutic protein production.
What is ‘pharming’ in biotechnological terms?
The production of recombinant therapeutic proteins in the milk of transgenic animals.
What are vectors in the context of recombinant protein production?
Plasmids that carry the gene of interest and facilitate its expression in a host.
What is the function of a selectable marker in a vector?
To allow for the identification of successfully transformed cells.
What is the purpose of the lac operon?
To control the expression of genes in bacterial systems.
What does IPTG do in bacterial expression systems?
Acts as an inducer to turn on the expression of genes downstream of the T7 promoter.
What is blue-white screening?
A method to identify recombinant clones based on color change in bacterial colonies.
What do restriction enzymes do?
They cut DNA at specific sequences to facilitate cloning.
What is the role of DNA ligase?
To join two DNA molecules together using energy to form a phosphodiester bond.
What are genomic DNA libraries?
Collections of cloned DNA that represent all genetic material of an organism.
What is the primary purpose of a cDNA library?
To represent the expressed genes within a particular tissue at a specific developmental stage.
What is a non-directional cloning strategy?
A cloning method that does not specify the orientation of the insert DNA in relation to the promoter.
What is a directional cloning strategy?
A cloning method that incorporates restriction sites in primers to ensure the insert is in a specific orientation.
What is the significance of polymerase chain reaction (PCR) in cloning?
To amplify the DNA of interest, generating sufficient quantities for cloning.
What does ‘transformation’ refer to in molecular biology?
The process of introducing recombinant DNA into host cells.
What is the function of phosphatase treatment in cloning?
To prevent self-ligation of the vector by removing the 5’ phosphate group.
What is the role of the SARS-CoV-2 spike protein in vaccine development?
It triggers an immune response when delivered via recombinant viral DNA.
