Bacterial_gen Flashcards
parts of plasmid define: ori(pBr322)
origin of replication for plasmid pBr322
parts of plasmid define: f1 ori
origin of replication specific to f1 cells
parts of plasmid define: KanR
kanamycin resistance gene used for selection
parts of plasmid define: bom (oriT)
necessary for conjugation
parts of plasmid define: rop
small protein that regulates plasmid DNA replication by modulation of transcription initiation of primer RNA precursor
parts of plasmid define: lacI/lacI promoter
repressor used for selectable expression of downstream genes
parts of plasmid define: lacO
operon required for transport/metabolism of lactose, used for inducible expression of genes of interest
parts of plasmid define: T7 terminator
transcription termination site to ensure desired transcript produced
parts of plasmid define: Restriction sites
handy for cloning and determining if your plasmid incorporated an insert
parts of plasmid define: Primer sites
handy for amplification and sequencing to see what you actually transfected in a large array
parts of plasmid define: AviTag-MPB-Ruby-His
Assorted tags that can be added to protein to purify easily
genetically tractable
able to take up a plasmid and express markers on those plasmids, as well as grow on media we are capable of making
suicide vector components
selectable marker, counter selectable marker, mutation of interest
why is suicide vector forced to integrate by homologous recombination? (2)
- Has a non-functional ori
2. Selection for selectable marker (antibiotic resistance)
why do we need a counter selectable marker on suicide vector?
Because we perform a second crossing over event, and need to select against the cells which don’t have a clean background by removing them from population (negative selection)
