Biotechnology and Recombinant DNA Chapter 9, pp 219-244
A) Recombinant DNA = joining of DNA from different sources
1)” tools” required
a) molecular scissors to cut out gene of interest = restriction
enzymes
(Table 9-3 p232)
blunt ends vs. sticky ends
b) vector to carry gene into recipient cell; includes plasmids,
viruses
2) process Fig 9-2, p 222
a) cut out desired gene from source
b) insert into appropriate vector e.g. plasmid
c) insert vector into recipient cell
d) select for transformed cells
3) preparation of cDNA = mRNA to DNA via reverse transcriptase
(no introns!) Fig 9-13, p232
4) uses of genetic engineering/biotechnology - see Table 9-1, p 220,
Table 9-2, p 221
B) Important techniques developed with biotechnology
1) Agarose gel electrophoresis
= separation of DNA pieces (fragments by size)
2) Polymerase Chain Reaction (PCR) = amplifies (makes many copies
of) pieces of DNA
3) DNA fingerprinting (what OJ has nightmares about!); also known
as RFLPs (riflips) for “restriction fragment length
polymorphisms.” Actually represents a way of looking at DNA
sequence homology (shows genetic relatedness).
see perspective 9-1, p228
4) Southern blotting Fig 9-8, p227
5) Colony blots Fig 9-7, p226
6) Microarray Fig 9-10, p227
7) DNA sequencing
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