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Recombinant DNA Technology

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Western University
Biochemistry 2280A
Derek Mc Lachlin

Brandl Lecture 5 Notes 11/16/2012 Termination of Translation  Requires o One of the three stop codons  No tRNAs for stop codons o Specific termination factors  Binding of release factor to the A site occurs during termination  Release factor adds water to the chain rather than another amino acid, which releases the peptide from the tRNA and terminates translation o Catalysis reaction Antibiotics and Translation  Many antibiotics block bacterial translation  Its intricate molecular interactions and importance make it a prime therapeutic target  See table 7.3 – do not memorize mechanisms but understand how they work  It is important to recognize that most tools of recombinant DNA technology use chemical quantities of the DNA o While I will present the DNA as a single line, there are in fact many identical molecules in the reaction o Q: In a DNA sequencing reaction, you generally add about 100 ng of DNA. Assuming the DNA is 3000 base pairs, how many molecules of template are added? o One base 10ir has a mass of about 600 Daltons (g/mole) A: 3 x 10  1 mole of a DNA of 3000 bp has a mass of: 3000 bp x 600 g/bp = 18 x 10 (approx. 2 x 10 ) 6 -7  1 mole – -7x 10 , 6 mole – -14 ng (1 x 10 )  X = 1 x 10 / 2 x 10 = 5 x 10  How many molecules? 5 x 10 -1x 6.02 x 10 (Avogadro’s number) = 3 x 10 10 Recombinant DNA Technology Genetic Engineering  The techniques which allow DNA fragments from different sources to be recombined to make new molecules with unique features Synthetic Biology  The construction of new biological parts and systems  The re-design of existing, natural systems for useful purposes  Plant/crop and dog breeding are examples of humans manipulated the DNA of other organisms  What is novel about recombinant DNA technology? Comparison of Classical Genetic Techniques and Recombinant DNA Technology  Classical genetics o Slow: limited by the breeding time of the organisms and change genetic events o Limited to breeding species  Recombinant DNA technology o Rapid: as quick as a few days in some organisms o No limitations Significance of Recombinant DNA Technology  Research o Recombinant DNA technology is a powerful tool in understanding cellular and molecular structure and function o Recombinant DNA technology was critical in genome sequencing projects  Biotechnology o The use of organisms to do work for man o Modern biotechnology impacts man
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