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Biology Genetic Engineering Gel Electrophoresis

Biology Genetic Engineering Gel Electrophoresis. Juliana Machado Isabella Rojas Gabriel Felipe Galvis. Genetic Engineering. Also called genetic modification .

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Biology Genetic Engineering Gel Electrophoresis

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  1. BiologyGeneticEngineeringGel Electrophoresis Juliana Machado Isabella Rojas Gabriel Felipe Galvis

  2. GeneticEngineering Alsocalledgeneticmodification. Isthe human manipulation of anorganismsgenome (hereditaryinformation of theorganism) usingmodern DNA technology. Itinvolvestheintroduction of DNA orsyntheticgenes into theorganism.

  3. History • Humanshavealteredthegenomes of speciesforthousands of yearsthrough artificial methods. Geneticengineeringconsideredthedirectmanipulation of DNA byhumans has onlyexisted in 1970s. • Paul Bergcreatedthefirsrrecombinant ( DNA sequencesthatresultfromthe use of laboratorymethods). Bycombiningthemonkey virus SV40 with lambda phage(another virus consisting of a head)

  4. Herbert and Stanley Cohen createdthefirsttransgenicorganismbyinsertingantibioticresistance genes intotheplasmid of anE. Colibacterium. • Rudolf Jaenischcreated a transgenic mouse byintroducing DNA intoitsembryo, makingitthefirsttransgenic animal. • In 1976, Genentech, thefirstengineeringcompanywasfoundedby Herbert Boyer and Robert Swanson.

  5. Applications Geneticengineering has applications in medicine, research, industry and agriculture. Medicine: Geneticengineering has beenusedto produce insulin, human growth hormones. Alsoitisusedtocreate animal modelswhich are non-human animals, usedduringtheresearch and investigation of human diseases. Geneticallymodifiedmice are themostcommongeneticallyengineered animal model.

  6. Research: Geneticengineeringisanimportanttoolfornatural scientists. Genes and othergeneticinformationfrom a widerange of organisms and information are transformedinto bacteria forstorage and modification, creatinggeneticallymodified bacteria in theprocess. Alsoorganismsare geneticallyengineeredtodiscoverthefunctions of certain genes. Thiscould be theeffectonthephenotype of theorganism, wherethe gene isexpressedorwhatother genes itinteractswith.

  7. Industry: Byengineering genes itispossibletocreate a biologicalfactorythat can produce proteins and enzymes. Bacteria and yeasthavebeenusedto produce medicines such as insulin, human growth and vaccines ( biologicalpreparationthatimprovesimmunityto a particular disease).

  8. Agriculture: Geneticengineeringisused in thecreation of geneticallyfood. Geneticallymodifiedorganismshavehadspecificchangesintroducedintotheir DNA bygeneticengineeringtechniques. Thesetechniques are much more precise, anexampleismutagenesis (mutationbreeding) whereanorganismisexposedtoradiationorchemicalstocreate a non- specificbutstablechange. Othertechniquesbywhichhumansmodifyorganismsinclude: selectivebreeding, plantbreedingand animal breeding. Sincegeneticallymodifiedfood has beenintroducedintosupermarkets, there has beenmuchcontroversy.

  9. Whatis Gel electrophoresis? When DNA manipulationbegan, DNA fragmentswereseparatedbygravity. In the 1970s, thepowerfultool of DNA electrophoresiswasdeveloped. Gel electrophoresis in chemistry: Isa methodusedtoseparateproteinsbycharge and size Gel electrophoresis in biochemistry: In molecular biologyitisusedtoseparate DNA and RNA fragmentsbylength, toestimatethesize of DNA and RNA.

  10. Electricityeffectsonmolecules • Nucleicacidsmolecules are separatedbyusing a electricfieldtomovethenegativelychargedmoleculesthroughanagarosematrix • Weneedtotakeintoaccountthatshortermoleculesmovefaster and migratelongerthanlongeronesbecauseshortermoleculesmigrate more easilythroughthepores of the gel. Thisprocessiscalledsieving. is a simple and convenienttechnique of separatingparticles of differentsize • In the case of proteinsthey are separatedbycharge in agarosebecuasethepores of the gel are toolargetosieveproteinsAlsoelectrophoresiscan be used in theseparation of nanoparticles.

  11. Agarose • Agaroseis a polysacharideobtainedfrom agar thatisusedfor a variety of scienceapplications, specially in gel electrophoresis. • Agaroseitisused in thematrix, whichplays a vital role in thisseparation.

  12. Agaroseused as a gel to be incluided in thematrix.

  13. Howdoes gel electrophoresiswork? -After DNA isextractedfromcellsorobtainedusing PCR (polymerasechainreaction), they are cutintomanyfragments of differentlenghtswithrestrictionenzymes (enzymethatcuts DNA).

  14. -DNA fragments are loadedintothewells of anagarosegel and placed in anelectrophoresis buffer filledtank. This gel whichis similar togelatinisformedforthe DNA fragmentsto be placed in here. -This gel is placed in thematrix and containselectrolytes, so thatanelectriccurrentconducted. -Positive and negativeelectrodes are connectedtotheoppositeends of the gel- (electricfieldisapplied). -Thenegativelycharged DNA fragments are drawntowardsthe positive terminal.

  15. The agar gel ishighlyporous • Shortest DNA fragments are abletomovequicklythroughthepores. • Larger DNA fragmentsmoveveryslowlybecauseitis more difficultforthemtopassthroughthepores.

  16. Applications of Gel electrophoresis • Estimation of thesize of DNA molecules. Example: restrictionenzyme (enzymethatcuts DNA) alsocalledrestrictionmapping of cloned DNA. • Analysis of PCR(polymerasechainreaction) productsis a scientifictechnique in molecular biologytoamplifyorincrease a single orfew copies of a piece of DNA acrossseveralorders, generatingthousandsormillions of copies of a particular DNA sequence. 8 PCR tubes, eachcontaining a reaction mixture

  17. Whatis DNA profiling? • DNA profilingalsocalled DNA fingerprinting, DNA testing, is a technique use byforensicscientiststodistinguishbetweenindividuals of thesamespeciesusingsamples of their DNA. • Althoughthemajority of our DNA isthesame, there are short piecescalledmicrosatellites . Theserepeatmany times in a persons DNA. Thisisthepattern of repeatswhichisdifferent in every individual. • Knowingthesemicrosatellites, DNA sequences are thebasisforthe DNA profiling.

  18. Stages of DNA profiling • STEP 1: Cells are brokendowntorelease DNA. If a smallamount of DNA isavailable, theamount can be increasedoramplifiedbyusingpolymerasechainreaction (PCR). • STEP 2: The DNA iscutintofragmentsusingrestrictionsenzymes as I explainedbefore. Eachrestrictionenzymecuts DNA at a specific base sequence. • Thesections of DNA which are cut are calledrestrictionfragments.

  19. STEP 3: In DNA profilingthe gel electrophoresisisinvolvedFragments are separatedonthebasisbyusing a processcalled gel electrophoresis. • DNA fragments are inserted in thewells and a electriccurrentisdeveloped. As I explainedbefore in gel electrophoresis. • DNA isnegativelycharged so itisattractedtothe positive part of the gel. • DNA isseparatedonthebasis of size. • A radioactiveisaddedwhich combines withthe DNA fragmentsto produce a image, thatcontainsthe copies of DNA bands. • STEP 4: Thepattern of thefragmentisanalyzed.

  20. USES OF DNA PROFILING • DNA profilingisusedtosolve medical problems. • Biological uses for DNA profiling: • Blood • Hair • Saliva • Bodytissuecells

  21. Bibliography • http://science.howstuffworks.com/dna-profiling1.htm • http://learn.genetics.utah.edu/search • http://www.sumanasinc.com/webcontent/animations/content/gelelectrophoresis.html • http://www.youtube.com/watch?v=QEG8dz7cbnY

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