TheMcGraw-HillCompaniesChapter 10GeneticEngineeringARevolutioninFoundationsinMolecularBiologyMicrobiologyEIGHTHEDITIOKathleenParkTalaroBarryChessCopyright The McGraw-Hill Companies, Inc. Permission required for reproduction or display
Chapter 10 Genetic Engineering: A Revolution in Molecular Biology Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display
Genetic Engineering: Basic knowledge is used to derive applied scienceor useful products Direct, deliberate modification of an organism'sgenome-BioengineeringBiotechnology - use of an organism'sbiochemical and metabolic pathways for industrialproduction2
2 Genetic Engineering • Basic knowledge is used to derive applied science or useful products • Direct, deliberate modification of an organism’s genome – Bioengineering • Biotechnology – use of an organism’s biochemical and metabolic pathways for industrial production
Tools and Techniques ofGenetic EngineeringPractical Properties ofDNAIntrinsic properties of DNA hold true even in a test tubeCopyright The McGraw -Hill Companies, Inc. Permission required for reproduction or dis play.DNA heated from 9o°C to95° C: the two strandsseparate.ThenucleotidesHeatingCoolingcan be identified, replicated,ortranscribed.Slowly cooling the DNAallows complementary(a)DNA heatingand cooling.DNAresponds to heat bydenaturing-losingnucleotides to hydrogenitshydrogenbondingand therebyseparating into itstwostrands.Whencooled,thetwostrandsrejoinatcomplementaryregions.Thetwostrandsbond and the DNA willneednotbefromthesameorganismsas long astheyhavematchingsitesregain double-stranded form3
3 Tools and Techniques of Genetic Engineering Practical Properties of DNA • Intrinsic properties of DNA hold true even in a test tube • DNA heated from 90°C to 95°C; the two strands separate. The nucleotides can be identified, replicated, or transcribed. • Slowly cooling the DNA allows complementary nucleotides to hydrogen bond and the DNA will regain double-stranded form Heating Cooling (a) DNA heating and cooling. DNA responds to heat by denaturing—losing its hydrogen bonding and thereby separating into its two strands. When cooled, the two strands rejoin at complementary regions. The two strands need not be from the same organisms as long as they have matching sites. Copyright © The McGraw -Hill Companies, Inc. Permission required for reproduction or display
EnzymesforDicing,Splicing,andReversingNucleicAcidsRestriction endonucleases - recognize specific sequencesof DNA and break phosphodiester bondsbetweenadjacent nucleotidesThe enzymes can be used to cleave DNA at desired sitesRecognize and clip the DNA at palindrome basesequencesUsed in the lab to cut DNA into smaller pieces -restrictionfragmentsCopyright The McGraw-HillCompanies, Inc. Permission required for reproduction or displayEcoRIHindlllEndonucleaseHaelllCuttingGAATTCGCTT.AGApatternCTTAAGT'TCGAACC4(b)Examplesofendonucleases,palindromes andcuttingpatterns.Thefirsttwo arestaggered cuts,andthethird isablunt cut
4 Enzymes for Dicing, Splicing, and Reversing Nucleic Acids Restriction endonucleases – recognize specific sequences of DNA and break phosphodiester bonds between adjacent nucleotides • The enzymes can be used to cleave DNA at desired sites • Recognize and clip the DNA at palindrome base sequences • Used in the lab to cut DNA into smaller pieces – restriction fragments (b) Examples of endonucleases, palindromes and cutting patterns. The first two are staggered cuts, and the third is a blunt cut. Cutting pattern Endonuclease G A A T T C C T T A A G EcoRI A A G C T T T T C G A A HindIII G G C C C C G G HaeIII Copyright © The McGraw -Hill Companies, Inc. Permission required for reproduction or display
MICRestrictionEndonucleasesawGAATTCDNADuplexCTTAAG国PlayPauseAudioTextRestrictionendonucleases areenzymesthat cleaveDNAatspecificnucleotidesequences.The sequencerecognizedis oftenfour to sixnucleotideslong.Forexample,therestrictionendonuclease EcoRI recognizesthesequence,GAATTCCopyright@TheMcGraw-Hill Companies,Inc