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Biotechnology and Genetic Engineering PBIO 450/550

Biotechnology and Genetic Engineering PBIO 450/550. Biotechnology terminology Common hosts in biotechnology research Transcription & Translation Prokaryotic gene organization & expression. Terminology.

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Biotechnology and Genetic Engineering PBIO 450/550

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  1. Biotechnology and Genetic EngineeringPBIO 450/550 Biotechnology terminology Common hosts in biotechnology research Transcription & Translation Prokaryotic gene organization & expression

  2. 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

  3. 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

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

  5. Common host organisms usedin molecular biotechnology • E. coli • Yeast (Saccharomyces cerevisiae) • Insect cell lines • Plant cell lines • Animal cell lines

  6. 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

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

  8. Chemical structure of DNA & RNA

  9. Chemical structure of dsDNA

  10. Chemical structure of dsDNA

  11. Prokaryotic gene expression

  12. Prokaryotic gene expression • Alternatively, see: • http://www.whfreeman.com/lodish4e/con_index.htm?99anm

  13. 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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