Bio 105 – Guide 06
http://www.ucalgary.ca/~einsiede/graphics/cartoon.gif
Biotechnology is a New Discipline
Chemists have had an "industrial", practical side
for the last 100 years. Manipulating molecules
through chemistry for practical purposes.
Until recently, Biologists were mainly observers of
living things. Where they could bring about
change, they had to do so within limitations. For
example, new traits were identified (or made by
mutation) and used. New combinations of genes
were only possible where breeding was possible.
Biotechnology is a New Discipline
Biologists are no longer limited by "natural
barriers"
•We can now cross species barriers and any
organism can be the recipient of any gene
from any other organism
•We can make any product, protein or RNA,
in large amounts - even if they normally exist
in very small amounts!!
Some History
1973 Stanley Cohen and Herbert Boyer – UC San
Francisco – first gene insertion
•The Asilomar Meeting
Safety Concerns
Ethical Concerns
•NIH – RAC
•June 7, 1987 – US. Patent Office allows one to
patent higher life forms – this made many advances
commercially attractive and greatly increased the
pace and practical results of research. The
commercialization of Biology
Enzyme tools are at the heart of
genetic engineering
•Nucleases
•Polymerases
•Ligases
Nucleases
A nuclease is an
enzyme capable
of cleaving the
phosphodiester
bonds between
the nucleotide
subunits of
nucleic acids.
• Exo
• Endo
–Non Specific
–Site Specific
Bacteriophage lambda Host Restriction
http://www.personal.psu.edu/rch8/workmg/Isolating_analyzing_genes_files/image007.png
http://www.science.siu.edu/microbiology/micr302/figure%209.12.JPG
Examples of Restriction Enzymes
Enzyme
Ava I
Bam HI
Bgl II
Eco RI
Eco RII
Hae III
Hha I
Hind III
Hpa I
Kpn I
Mbo I
Mbo I
Pst I
Sma I
SstI
Sal I
Taq I
Xma I
Organism from which
derived
Anabaena variabilis
Bacillus amyloliquefaciens
Bacillus globigii
Escherichia coli RY 13
Escherichia coli R245
Haemophilus aegyptius
Haemophilus haemolyticus
Haemophilus inflenzae Rd
Haemophilus parainflenzae
Klebsiella pneumoniae
Moraxella bovis
Moraxella bovis
Providencia stuartii
Serratia marcescens
Streptomyces stanford
Streptomyces albus G
Thermophilus aquaticus
Xanthamonas malvacearum
Target sequence
(cut at *)
5' -->3'
C* C/T C G A/G G
G* G A T C C
A* G A T C T
G* A A T T C
* C C A/T G G
GG*CC
GCG*C
A* A G C T T
GTT*A A C
GGTA C*C
*G A T C
*G A T C
CTGCA *G
CCC*GGG
GA GCT*C
G*TCGA C
T*CGA
C*CCGGG
For average DNA, the size of the Restriction
Endonuclease recognition site determines the
frequency of the sensitive sites
Recognition
Site of (no. of bases)
4
6
8
Cutting Average
every (bp)
256
4096
65536
http://www.mun.ca/biology/scarr/Lab_5_gel_1999.gif
DNA polymerase is an enzyme (the
suffix -ase is used to identify
enzymes) that helps catalyze the
polymerization of DNA bases
(deoxyribonucleotides) into a DNA
strand. DNA polymerases are best
known for their feedback role in
DNA replication, in which the
polymerase "reads" an intact DNA
strand as a template and uses it to
synthesize the new strand.
DNA ligase is a specific type of enzyme, that facilitates the
joining of DNA strands together
http://universe-review.ca/I10-71-plasmid.jpg
Cells with the
intact pUC19 can
catabolize X-gal
generating blue
colored colonies,
while cells
containing the
recombinant
plasmid remain
white.
http://photoscience.la.asu.edu/photosyn/courses/BIO_343/lab/plasmid.gif
http://www.edvotek.com/images/300.jpg
Using Reverse
Transcriptase to
Clone Genes
http://www.mun.ca/biology/desmid/brian/BIO
L2060/BIOL2060-20/2031.jpg
http://faculty.ircc.edu/faculty/tfischer/images/transformation.jpg
The process in which a bacteriophage
(virus) transfers bacterial genes from
one bacterium to another is called:
a. conjugation
b. transposition
c. recombination
d. transduction
e. transformation
http://faculty.ircc.edu/faculty/tfischer/images/generalized%20transduction.jpg
http://biology200.gsu.ed
u/houghton/4564%20'04
/figures/lecture%203/co
njugation.jpg
http://upload.wikimedia.org/wikipedia/en/thumb/2/29/BacterConjugation.png/350px-BacterConjugation.png
The End