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|00010|00020|00030|00040|00050|01000|02000|03000|04000|05000|06000|07000|08000|09000|10000|11000|&ns=0

Central Dogma of Biology transcription translation DNA RNA Protein $$$ reverse transcription Molecular Biotechnology DNA replication

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_index.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