PPT-A personal history of restriction enzymes

Richard J Roberts New England Biolabs CSHL 2013 AHA Danna K Nathans D 1971 Specific cleavage of simian virus 40 DNA by restriction endonuclease of Hemophilus influenza Proc Natl Acad Sci USA 68 29132917 . Restriction Enzymes. General Genetics. Objectives:. Introduce the students to digest genomic DNA by restriction . endonucleases. .. Observe the results of digestion on . agarose. gel electrophoresis.. GENETIC. . ENGINEERING. Genetic engineering is the manipulation of genetic materials which can be introduced in the host organisms and thus change the phenotype of the host organism.. IMPORTANCE. OF ENZYMES. Enzymes are biological catalysts that:. . I. ncrease. . reaction rates . by lowering the amount of energy needed for the reaction to occur. 100-1000x . faster. !!!. Activation Energy. Objectives. -Understand how Restriction Enzymes digest DNA.. -Know how to construct a . pAMP. . (plasmid) or gel.. -Given the size of fragments, gel, know how to construct a restriction map.. -Given a restriction map know how to construct a gel.. BIOTECHNOLOGY. What. . is. . biotechnology. ? . Biotechnology. = . bios. (. life. ) + logos (. study. . of). Literally. ‘. the. . study. of . tools. . from. living . things. ’. What. . What are restriction endonucleases (REs)?. How can REs be used to identify DNA molecules?. How can I find RE recognition sites in the . MET. plasmids? . Restriction endonucleases are part of a bacterium’s defense against invaders. Enzymes“. „. Sequence. . specific. . recognition. . and. . engineering. “. Alfred Pingoud. CSHL. Oct. . 19-21 2013. roberts@neb.com. . Sa 03.08.2013 . Outline of . talks. Alfred . Pingoud (25 . Restriction enzymes. Polymerase enzyme. Strand-displacing polymerases. Helicase enzymes. Discovery and Obtainability:. Most enzymes are proteins discovered in cells.. But . DNAzymes. were . descovered. are. . working. ?. Restriction. Enzymes. Natural . occurrence. . of. . restriction. . enzymes. . is. in . bacteria. . for. . protecting. . them. . against. a . virus. . attack. . by. . : An enzymes that breaks down nucleic acid molecule by breaking the phosphodiester bonds at end 3. / . or 5. /.. Endonucleases. : . An Enzyme that hydrolyzes internal phosphodiester bonds in a polynucleotide chain or nucleic acid molecules.. the isolation of the restriction . endonuclease. from. E. coli K (. Meselson. & Yuan 1968). . It was evident . that the restriction . endonucleases. from . E. coli K . and . E. coli B were important examples of proteins. Restriction Modification System. Cutting DNA molecules. . Before 1970s, there was no method of cleaving DNA at the specific points.. All the available methods for fragmenting DNA were non-specific.. Presented . by. Ms. . P. . . H. . Giri. Department of Microbiology. Deogiri . College, Aurangabad. B.Sc. T. Y. . Semester VI. Paper No. XIX. Recombinant DNA Technology. Ms. . Priyanka. H. . Giri. Steps in RDT:. by. Prof. . Meena. . Nagawanshi. Professor . Department of . Zoology. Deogiri . College, Aurangabad. Enzymes. Enzymes are proteins. . biological catalysts .  help drive biochemical reactions. Enzyme names end with an .