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MCB 720: Molecular Biology •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 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 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 5Protein sorting http://bcs.whfreeman.com/lodish5e/pages/bcsmain.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 transcription RNA reverse transcription DNA replication translation Protein $$$ 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