MCDB 3135 Study Guide - Quiz Guide: Expression Vector, Nucleic Acid Thermodynamics, Antimicrobial Resistance
28 Mar 2023
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Topics for Quiz 5 (Technology)
Questions in the quiz will deal with topics discussed on the slides. No need to read the book, but
keep in mind that looking into the book does not hurt …!
- Make yourself familiar with chromatin structure. What is a gene, transcription, exon,
intron, etc. This will follow us during the entire class.
- What is PCR, and what can you do with it. How does the amplification of a gene works
(primers, DNA melting temperature, annealing of primers, why TAC polymerase, not just a
eucaryotic or bacterial polymerase which might be more efficient?
- You have a PCR product. What are you do with this next? Why would you engineer
restriction sites at each end of the chain? How do you insert your amplified piece into an
expression vector? What is an expression vector?
- How do you design primers for a gene? What has to be considered regarding sense and
antisense strand?
- What are restriction enzymes? What do they recognize, and where do they cleave a DNA
strand (exo-, or endonuclease) ??
- What is EcoR1, and what does the name mean?
- What is the difference between blunt and sticky end? How would you make sure your gene
inserts into your vector with the correct polarity? What is an ORF?
- How useful are restriction enzymes, and what is the chance of finding a random site on a
human genome, as compared to CRISPR Cas9?
- What is the function of RecA?
- How is an expression vector designed? What are common features of these plasmids
(antibiotic resistance gene, polylinker, promoter, operator, origin of replication)? Where
will you insert your gene? What is controlling the level of your gene expression?
- What is the origin of replication do? How could this influence the amount of protein you
will synthetize in a cell.
- Why do we usually use an antibiotic resistance gene on expressed plasmids (selection) ?
