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  • Introduction Third Draft

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    jerichopaolos591
    17 vues4 pages
    RDNA technology is being used to make synthetic insulin through yeast. This is because yeast is eukaryotic and is processed almost the same way human proinsulin is made. The objective of this experiment is to open a new way to produce insulin, which will ultimately make it available, abundant, and inexpensive for all.

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    RDNA technology is being used to make synthetic insulin through yeast. This is because yeast is eukaryotic and is processed almost the same way human proinsulin is made. The objective of this experiment is to open a new way to produce insulin, which will ultimately make it available, abundant, and inexpensive for all.

    Droits d'auteur :

    © All Rights Reserved
    Téléchargez comme PDF, TXT ou lisez en ligne sur Scribd
    Signaler comme contenu inapproprié
    17 vues4 pages

    Introduction Third Draft

    Transféré par

    jerichopaolos591
    RDNA technology is being used to make synthetic insulin through yeast. This is because yeast is eukaryotic and is processed almost the same way human proinsulin is made. The objective of this experiment is to open a new way to produce insulin, which will ultimately make it available, abundant, and inexpensive for all.

    Droits d'auteur :

    © All Rights Reserved
    Téléchargez comme PDF, TXT ou lisez en ligne sur Scribd
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    sur 4

    Introduction

    Theproductionofsyntheticinsulinmarkedamilestoneinthebiotechindustry.Through
    theuseofrDNAtechnology,millionsofpeopleareabletoregulatetheirbloodsugarlevels
    fromatimeofinsulinshortageandpreposterousprices.Preproinsulinisbeingusedtomake
    syntheticinsulinthroughyeast.Thisisbecauseyeastiseukaryoticandisprocessedalmostthe
    samewayhumanproinsulinismade.Lookingbacktoamoreprimitiveprocess,insulinwas
    oncemadethroughthethousandsofpoundsofpigpancreas,onlytocreate8ouncesof
    purifiedinsulin(GriffithsAJF,1997).Today,Insulinismadein1.38millionacrefacilities
    insidefivethousand,50,000fermentorsdesignedtomakeinsulincrystalsbythemetrictona
    day(Gebel,2013).Thissurgeinthebiotechindustrycreatesageneroussupplyofinsulin,as
    wellashavingasalesprofitof$16.7billiondollars,anditsexpectedtogrowwiththe
    introductionofgenericinsulinchoices.Thepatentprotectiononinsuliniscomingtoanend,
    andwiththeinsulinmarketopeningup,waysofprovidingafasterwaytoproducehigh
    quality
    insulinwillbesought.Genentech,acompanythatcreatedthefirstrDNAproductintheworld,
    hascreatedgeneticvariationstotreatexistingdiseasessuchascancer,diabetes,andothers.
    Withnewbacteriafrontsonthemarket,biotechindustriescanexpandevenfurthertocreate
    moreversatiletreatmentsasopposedtobeinglimitedtoE.Coliastheonlychoiceofbacterial
    reproductionofhumanhormones(Kaminoka,2011).Theobjectiveofthisexperimentisto
    openanewwaytoproduceinsulin,whichwillultimatelymakeitavailable,abundant,and
    inexpensiveforall.Thesefindingsshowthat[findings]and[conclusion].


    MethodsandMaterials
    LBAgar/AmpPetriDishes
    E.ColicanbecultivatedinLBAgarplates,andE.Colimodifiedtohaveampicillin
    resistancecangrowbecultivatedinLBAgar/Ampplates.Weight3.5gofLBAgarpowder
    withanelectronicscale,andthenmixthepowderwith250mlofdH2O.Then,microwaveuntil
    yellowandtranslucent.Afterthesolutionwasyellowandtranslucent,itwaspouredintopetri
    dishesuntilitwasusedup,whichmadeabout10plates.Thesameprocesswasusedtomake
    LBAgar/Ampplates,butafterthesolutionwasmicrowaveduntiltranslucentyellow,ithasto
    becooled.Oncecooled,100uLofampicillinresistancewasaddedintothesolution.After
    mixed,thesolutionwaspouredintopetridishesandlefttheretocoolandsolidify.Afterthegel
    wassolid,itwasstoredbysealingwithparafilmandputintotherefrigerator.
    TransformingE.Coli
    E.Colicanbetransformedtoproduceinsulinatafasterrateoriginalmethods.Twotubes
    werelabeled+insulinandinsulin.250uLoftransformationsolutionwereaddedintobothtues
    andthenplacedonice.FourcoloniesofE.Coli,byuseofasterileloop,aretransferredinto
    eachtube,usinganewloopforeachtube.10uLofInsulinDNAistransferredintothe+insulin
    tube.Bothtubesareincubatedonicefortenminutes.Platesarelabeled:+Insulin
    LBAgar/Amp,InsulinLBAgar,andInsulinLBAgar/Amp.Afteraquicktransferfromice,
    incubatedtubeswereplacedintoa42degreeCelsiuswaterbathfor50seconds,andthen

    transferthembackoniceforanothertwominutes.Afterwards,250uLofLBnutrientbroth
    wereintoeachtube.100uLoftheappropriatetransformationwerethentransferredtoits
    designatedplate.Byuseofasterileloop,thesolutionwasspread,parafilmed,andstored
    upsidedowninside37degreeCelsiusincubator.
    DNAPurification
    [Notfinished,noresultsyet]

    References:

    Gebel,E.(2013,July1).MakingInsulin.RetrievedNovember18,2014,
    fromhttp://www.diabetesforecast.org/2013/jul/making
    insulin.html
    Kamionka,M.(2011).EngineeringofTherapeuticProteinsProductioninEscherichiacoli.
    Current

    PharmaceuticalBiotechnology,12(2),268274.doi:10.2174/138920111794295693

    Howinsulinismadeusingyeast::DNALearningCenter.(n.d.).RetrievedNovember18,

    2014,fromhttp://www.dnalc.org/view/15929
    How
    insulin
    is
    made
    using
    yeast.html

    GriffithsAJF,GelbartWM,MillerJH,etal.ModernGeneticAnalysis.NewYork:W.H.
    Freeman

    1999.ExpressingEukaryoticGenesinBacteria.Availablefrom:

    http://www.ncbi.nlm.nih.gov/books/NBK21359/

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