Recombinant DNA versus Molecular cloning
recombinant DNA = construction methodology
(biochemistry)
molecular cloning = realisation of the power of
recombinant DNA technology
Two fundamental needs of the molecular biologist
1) separate an heterogenous mixture of DNA molecules
into separate entities
2) amplify the isolated DNA molecules
(extensive duplication)
Cloning of cells (bacteria, yeast, plant, mammal, etc.)
E. coli (Escherichia coli) as workhorse
single cells => colony on solid substrate
cells from colony into tube with growth medium
“genetically pure” : isogenic
Cloning of DNA :
1) needs capability to replicate
2) equal partitioning to the daughter cells
=> use of a vector : plasmid or other extrachromosomal element
Cloning in viruses : packaging
in vitro packaging
Introduction into host cells : adapt cells to uptake
transformation versus transfection
versus infection
versus transduction
Summary :
cloning of cells
viruses
=> colony
=> plaque (lysis zone)