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20 Cards in this Set

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  • Back
restriction enzymes
DNA cutting ezymes; cuts phosphodiester bond that joins adjacent nucleotides
bind to, recognize, cut DNA w/ specific sequences of bases, usualy 4 or 6 nucleotides
plasmid DNA
circular form of DNA
primarily found in bacteria; extrachromosomal b/c present in bacerial cytoplasm and bacterial chromosome; self replicating
Cohen and Boyer
cut plasmid and joined together fragments from each using DNA ligase to creat new, hybrid

boyer: cutting and sticking
cohen: insertion and replication of plasmid DNA
=sticking DNA together

sticky ends b/c can form base pairing H bonds, seals ends

1. 2 pieces of DNA w/ complimentary ends
2. H bonds form b/w complimentary bases
3. ends are sealed by ligase
how to get piece of DNA into a cell
-chemical treatments
-lipid based systems
-particle bombardment
carrier for foreign/target DNA
pieces of DNA that can accept,carry, replicate other pieces of DNA
restriction mapping
cloned DNA separated and visualized showing sites taht are cut by specific restriction enzymes
gel electrophoresis
separationg of cloned fragmens on gel according to size
migrates from - to +
smaller pieces move faster (toward + end), travel furthest
southern blot test
1. gel electrophoresis: separate fragments by size (see which fragments contain novel gene)
2.treat with alkaline solution to denature DNA, transfer fragments to nitrocellulose (blotting)
3.hybridize base pairs using piece of labeled DNA as probe
single stranded fragments of DNA or RNA w/ complementary ccode for specific sequence of bases
cuting DNA with number of restriction enzymes and getting fragments of DNA which vary in size from individual to individual
cutting DNA at specific sites, so fragments collected are various sizes as result of individual's number of repeats in specific introns. can use PCR to amplify amount of DNA in sample
DNA fingerprints
extract DNA, cut using restriction enzymes, separate fragments by size on a gel, transfer to nitrocellulose (southern blot), hybridize using a specific radioactive VNTR
=making lots of copies of a piece of DNA
1.separate strand of target by denaturing (heating)
2.add 2 specific primers that recognize/hybridize with either end of target and go in converging directions
3.again heat to denature the 4 strands
4.lower temp to allow primers to anneal (hybridize) to all 4 strands
5.raise temp again to synthesize polymerase
francis collins
human genome project
craig venter
used public info to push public effort of human genome project
criticized by many, watson
what mRNA tells us
what genes being expressed at what levels
level tells us which proteins being produced and functioning
complementary DNA
stable, can be measured quantitatively
=all the protein contained w/in a cell or organism
reflect state of a biological system
bovine heart proteins
separate proteins according to size and acidity