30 Questions
What is the process of making cells acquire and express a new piece of foreign genetic material called?
Transformation
Which term refers to the organism that has had its genome artificially altered by introducing foreign DNA sequences?
Transgene
What is the term for a DNA molecule used to carry a foreign DNA segment into a host cell?
Recombinant plasmid
Which process involves introducing recombinant plasmid to animal eukaryotic cells via artificial means?
Transfection
What is the term used for a cell that has taken up the recombinant plasmid and acquired a new trait?
Transformed cell
What is the term for the protein produced by the inserted foreign DNA?
Recombinant protein
What is the function of the origin of replication in plasmids?
Helps bacteria make copies of the plasmid
What is the purpose of antibiotic resistance in genetically engineered bacteria?
To allow transformed bacteria to survive in the presence of antibiotics
Where does the GFP chromophore reside within the GFP protein structure?
At the center of the protein
What is the primary function of plasmids in genetic engineering?
To provide a means for scientists to modify DNA
How is the GFP chromophore described in terms of its structure?
Comprised of three adjacent amino acids
What makes the GFP molecule notable in scientific circles?
Its ability to fluoresce under UV light
What enables cells with the plasmid to grow in culture media with ampicillin?
Secretion of Beta-lactamase
What prevents the GFP gene from being transcribed in the absence of arabinose?
Blockage of RNA polymerase by AraC
What is the role of arabinose in the system described?
Acts as a switch for gene expression
Why do cells without the plasmid fail to grow on plates with ampicillin?
They are unable to break down ampicillin
What happens if cells with the plasmid are grown in culture media without arabinose?
They will not produce GFP
Which protein plays a role in switching on the transcription of the GFP gene?
AraC
What is the first step in producing a protein of interest?
Identify a gene for the protein
What happens after the gene for a protein is identified?
Put the gene into bacteria
What is the role of bacteria in producing a protein of interest?
Transcribe and translate the gene
Which step involves making a plasmid with the gene for production?
Purify the protein of interest
What happens after growing lots of bacteria in the protein production process?
Purify the protein of interest
In genetic engineering using plasmids, what does bacterial transformation involve?
Put the gene into bacteria
What is the purpose of Hydrophobic Interaction Chromatography (HIC) in GFP purification?
To isolate the hydrophobic protein of interest, such as GFP.
What is the purpose of washing less hydrophobic proteins from the HIC column?
To remove the hydrophobic proteins from the column matrix.
What is the function of the Elution buffer in HIC chromatography?
To elute the target protein from the column.
Why is it important to discard the liquid broth supernatant before resuspending the pellet with TE buffer?
To avoid contaminating the sample with other proteins.
Which step ensures that only the hydrophobic protein of interest remains bound to the HIC column?
Washing with medium salt buffer.
What is the primary characteristic that enables GFP to be isolated effectively using Hydrophobic Interaction Chromatography?
Its hydrophobic nature.
Explore the methods for producing proteins of interest, such as modified mosquitoes for disease resistance, drug production like insulin and vaccines, and gene insertion into bacteria like E. coli. Learn about identifying genes and the process of protein production.
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