RECOMBINANT DNA TECHNOLOGY

What is different between these 2 sequences?

GGAATTCCTAGCAAT

CCTTAAGGATCGTTA

CTACGTGAGGAATTC

GATGCACTCCTTAAG

Only color & order of bases! The actual chemical nucleotides are the

SAME, even though the blue is sequence from US (eukaryotes) and the green is from bacteria (prokaryotes)!

So, DNA in plants, bacteria and us is chemically the SAME!!! That means, bacteria can “read” our DNA! (transcribe & translate our genes!)

Because of this, we can put our genes (or plant genes) in bacteria- this is what Recombinant DNA technology does!

TOOLS OF RECOMBINANT DNA Plasmid : a small circular DNA molecule

that is not part of the bacterial chromosome; replicates independently

Restriction Enzyme: Molecular scissors that cut DNA at a specific nucleotide sequenceEx.: EcoRI CATCGAATTCAC

GTAGCTTAAGTG

TOOLS, CONT. DNA Ligase: An enzyme that joins 2 DNA

fragments together (the “glue gun”).

Escherichia coli strains: Weakened strains of bacteria that are used to grow the plasmids.

Transformation: Process used to put the

plasmid back into the bacteria

HOW IS IT DONE?

ANIMATIONS: http://present.smith.udel.edu/biotech/rD

NA.html

WHY PUT A HUMAN OR PLANT GENE IN BACTERIA? Make lots of copies of the sequence

More of an issue before the human genome project Mutate or change 1 human gene separate

from the other 28,000 genes…. Make lots of a human protein cheap & easily!

(ex. is human insulin)