lecture 4 Flashcards
recombinant dna technology
copy, clone and manipulate RNA
purpose of restriction enzymes
cut dna
purpose of dna coning vectors
accept dna
molecular cloning steps
- isolate and purify target gene
- cut vector
- ligate/ glue gene into vector
- put vector/ gene into bacteria
properties of cloning vector:
should be fairly small dna molecules why?
get into bacteria
properties of cloning vector:
should have an origin of replication why?
make copies
properties of cloning vector:
should have at least one selectable marker
example
why
-color change or antibiotic resistance
-detection
properties of cloning vector:
should have at least one unique endonuclease recognition site
why
allow you to cut open vector
Colin bacteriocin gene removed! why
b/c would kill the bacteria
multi copy plasmids
bacteria can make many copies
Cut Vector and Gene of Interest with same restriction enzyme
* What must you keep in mind when doing this?
- only cut vector once
- never cut critical genes
Combine Vector and Insert with what enzyme to join ends?
ligase gives
What techniques are used to detect insert?
- run a dna gel
- insertional inactivation
- reporter genes
reporter
color change
since lambda is bacteriophage what does that tell us
that it infects bacteria
***Tumor-inducing
argrobacterium tumerfactons
(bacteria that infects plans)