E. coli

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14 Δεκ 2012 (πριν από 4 χρόνια και 9 μήνες)

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Biotechnology and Genetic Engineering

PBIO 450/550


Biotechnology terminology


Common hosts in biotechnology research


Transcription & Translation


Prokaryotic gene organization & expression

Terminology


Biotechnology
-
production of goods and services using
biological organisms, systems, and processes


Recombinant DNA technology
-
a set of techniques for
manipulating DNA, including: the identification and cloning of
genes; the study of the expression of cloned genes; and the
production of large quantities of gene product


Genetic engineering
-
the process of transferring DNA from one
organism into another that results in a genetic modification


Molecular biotechnology
-
rDNA technology + biotechnology

Table 1.1 Selected developments in the
history of molecular biotechnology

1944
-
Avery, MacLeod & McCarty determine DNA is the genetic material

1953
-
Watson & Crick determine the structure of DNA

1970
-
first restriction endonuclease isolated

1973
-
Boyer & Cohen establish recombinant DNA technology

1976
-
DNA sequencing techniques developed

1980
-
U.S. Supreme Court rules that genetically modified micro
-
organisms can
be patented

1981
-
first DNA synthesizers sold

1988
-
PCR method published

1990
-
Human genome project initiated

1996
-
Complete DNA sequence of a eukaryote (yeast) determined

1997
-
Nuclear cloning of a mammal (a sheep named Dolly)

2000
-
Arabidopsis

genome sequenced

2001
-
Human genome sequenced

2002
-
Complete human gene microarrays (gene chips) available


Fig. 1.2 Many scientific disciplines contribute to
molecular biotechnology, which generates a
wide range of commercial products

Common host organisms used

in molecular biotechnology


E. coli


Yeast (
Saccharomyces cerevisiae
)


Insect cell lines


Plant cell lines


Animal cell lines

Review protein secretion and protein targeting


Signal peptide sequences


Consider gram negative
vs. gram positive bacteria


Consider eukaryotic cells


In eukaryotic cells, short
peptide sequences (or
other modifications) tell a
protein where to go


See MCB Chapter 5
-
Protein sorting
http://bcs.whfreeman.com/lodish5e/pages/bcs
-
main.asp?v=category&s=00010&n=05000&i=05010.01&o=|00510|00610
|00520|00530|00540|00560|00570|00590|00600|00700|00710|0001
0|00020|00030|00040|00050|01000|02000|03000|04000|05000|060
00|07000|08000|09000|10000|11000|&ns=0

Central Dogma of Biology

DNA



RNA



Protein

transcription

translation

reverse

transcription

DNA

replication

$$$

Molecular

Biotechnology

Chemical structure of DNA & RNA

Chemical structure of dsDNA

Chemical structure of dsDNA

Prokaryotic gene expression

Prokaryotic gene expression


Alternatively, see:


http://www.whfreeman
.com/lodish4e/con_ind
ex.htm?99anm

In prokaryotes, RNA polymerase binds to the
-
10 and
-
35 regions of the promoter relative to
the start site of transcription (+1)

promoter

operator