recombination Flashcards
molecular scissors are
restriction endonucleases
molecular glue is
dna ligase
what are restriction enzymes
Cut up DNA
how can a bacterias own dna be protected from restriction enzymes
dna is methylated
restriction enzymes can produce cleaved dna with what 2 type of ends
sticky and blunt
sticky ends have what
over hangs
restriction enzymes cut where
recognition sites
recognition sites are often what type of sequences
palindromic
how long are recognition sites usually
4 or 6
restriction enzymes with the same recognition sites are called what
isoschizomers
What is needed for ligase to join DNA
5’ phosphate end
how can vectors be stopped from religating?
Use CIP to remove the end 5’ phosphate group and the phospho group needs to be provided by the insert
what are vectors
vehicles used to transfer foreign genetic material to a host cell
most cloning vectors are what?
plasmid or viral chromosomes
What 3 things must a cloning vector have?
- Origin of replication to direct self replication
- Dominant selectable marker, usually confers drug resistance on the host cell
- Unique restriction endonuclease cleavage site (one EcoRI site in a plasmid)
