Biotechnology and Genetic Engineering PBIO 450550 Biotechnology terminology
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-r. DNA technology + biotechnology
Table 1. 1 Selected developments in the history of molecular biotechnology 1944 -Avery, Mac. Leod & Mc. Carty 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/lodish 5 e/pages/bcsmain. asp? v=category&s=00010&n=05000&i=05010. 01&o=|00510|00610 |00520|00530|00540|00560|00570|00590|00600|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 transcription RNA reverse transcription DNA replication translation Protein $$$ Molecular Biotechnology
Chemical structure of DNA & RNA
Chemical structure of ds. DNA
Chemical structure of ds. DNA
Prokaryotic gene expression
Prokaryotic gene expression • Alternatively, see: • http: //www. whfreeman. com/lodish 4 e/con_ind ex. htm? 99 anm
In prokaryotes, RNA polymerase binds to the 10 and -35 regions of the promoter relative to the start site of transcription (+1) promoter operator
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