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