cloning vectors
polylinker
-DNA segment with several unique sites for restriction endonucleases located next to each other -Cutting plasmids with one of the restriction enzymes does not disrupt any of the essential features of the vector
plasmid vectors
-are about 1.2-3kb -contains 3 things: a replication origin sequence; a gene that allows selection; exogenous DNA can be inserted into the bracketed region
List 2 disadvantages of using plasmid cloning vectors
-cannot accept large fragments -standard methods of transformation are inefficient
list the 2 methods for transforming bacteria
-electroporation -CaCl2 treatment with heat shock
list the four common properties of cloning vectors
-have replication origin -contain a genetic marker (usually dominant) for selection -unique restriction sites to facilitate cloning of inserted DNA n(polylinker) -minimum amount of non-essential DNA to optimize cloning
transforming bacteria
-purpose is to insert the newly created plasmid into a bacterial host cell so that it can be replicated
yeast artificial chromosomes
-purpose: cloning vehicles that grow in eukaryotic cell hosts as eukaryotic chromosomes -can clone very large inserts of DNA
list the 5 advantages of using plasmid vectors
1- small size (easy to manipulate and isolate) 2- circular (more stable) 3-replication independent of host cell 4- several copies may be present (facilitates replication) 5- frequently have antibody resistance (detection easy)
bacteriophage
20+kb Foreign DNA is inserted into the phage head. Once genetic info is inserted, the tail will assemble to infect host cells
cosmid vectors
50kb, phage vector that can be circularized to replicate independently in a host or packaged for transduction
What are cloning vectors
DNA molecules that can be used to transport DNA sequences between biological hosts
list the 3 advantages of using cosmid vectors for cloning
Strong selection for cloning of large inserts Infection process rather than transformation for entry of chimeric DNA into E. coli host means greater transformation efficiency Maintain Cosmids as phage particles in solution
therapeutic cloning
The production of embryonic stem cells for use in replacing or repairing damaged tissues or organs
reproductive cloning
Using a somatic cell from a multicellular organism to make one or more genetically identical individuals.
expression vectors
contain elements to facilitate protein expression
shuttle vectors
optimized for expressing genes in different domains clone and amplify DNA in a bacteria
how do you know which is a recombinant vector
put the vectors in antibiotic medium, the vector that survives is recombinant because it is resistant to antibiotics