Name: recombinant dna in cloning and gmos
Uploaded: 2026-08-26T08:20:51.528Z
Duration: 1 min 34 s
Channel: JoVE

- [Instructeur] Recombinant DNA is DNA van verschillende bronnen, meestal verschillende soorten die worden gecombineerd in het laboratorium voor w&#x133;dverspreid gebruik in klinisch en wetenschappelijk onderzoek.
 Recombinant DNA bestaat gewoonlijk uit een gen van belang, hier insuline, uit een donor-organisme dat in een vector wordt ingebracht, waardoor DNA van een ander organisme zichzelf repliceert, zoals een virus of een plasmide, een kleine cirkelvormig stukje bacterieel DNA.
 Het DNA van elke bron wordt eerst door restrictie-endonuclease gesneden, enzymen die het DNA op een bepaalde sequentie splitsen.
Het donor-dna wordt aan de uiteinden van het vector-DNA verbonden met de hulp van het enzym DNA-ligase.
 Het recombinant DNA wordt dan ingebracht in een gastorganisme, een bacterie of gist, waar het gerepliceerd wordt, en zo vele kopie&#xEB;n genereert van het betreffende gen.
 Het gen kan dan gemakkelijker bestudeerd worden, of, zoals in het geval van insuline, vertaald worden in een prote&#xEF;ne door de cellulaire machine van de gastheer.

recombinant dna

Recombinant DNA Technology

<h4>Overzicht</h4>



Wetenschappers cre&#xEB;ren recombinant DNA door DNA uit verschillende bronnen &#x2013; vaak andere soorten &#x2013; in het laboratorium te combineren. Met DNA-klonering kunnen onderzoekers specifieke genen bestuderen door ze in gemakkelijk te manipuleren cellen, zoals bacteri&#xEB;n, te plaatsen. Organismen die recombinant DNA bevatten, staan bekend als genetisch gemodificeerde organismen (GGO&#x27;s). Recombinant-DNA-technologie produceert organismen met nieuwe genen die de wetenschap, de geneeskunde en de landbouw ten goede kunnen komen.



<h4>Hoe cre&#xEB;ren wetenschappers recombinant DNA?</h4>



Het cre&#xEB;ren van recombinant DNA omvat het inbrengen van een interessant gen in een vector &#x2013; een voertuig dat vreemd DNA naar gastheercellen transporteert voor DNA-replicatie en eiwitexpressie. De meest gebruikte kloonvectoren zijn plasmiden, kleine cirkelvormige stukjes DNA die zich onafhankelijk van het chromosomale DNA van de gastheer repliceren.

Om recombinant DNA te cre&#xEB;ren, worden zowel het donor-DNA, inclusief het betreffende gen, als de vector geknipt op specifieke nucleotidesequenties &#x2013; restrictieplaatsen genoemd &#x2013; met behulp van restrictie-enzymen. Het enzym DNA-ligase sluit de suiker-fosfaat-ruggengraat af waar het gen van belang en het plasmide met elkaar verbinden.

Het resultaat is een recombinant DNA-molecuul dat bestaat uit een vector met een ge&#xEF;ntegreerd stukje donor-DNA, een zogenaamde insert. Een wetenschapper kan dit hybride DNA-molecuul vervolgens in een gastheerorganisme introduceren &#x2013; meestal bacteri&#xEB;n of gist &#x2013; waar het zich gemakkelijk en snel vermenigvuldigt. Hierdoor ontstaan er veel kopie&#xEB;n van het betreffende gen, wat nodig is voor wetenschappelijk onderzoek en andere toepassingen. Het gen kan ook worden getranscribeerd en vertaald in het gewenste eiwit &#x2013; zoals humane insuline &#x2013; door gebruik te maken van de cellulaire machinerie van de gastheer.

Het cre&#xEB;ren van recombinant DNA is een onvolmaakt proces en er komen vaak fouten voor. De vector kan bijvoorbeeld sluiten zonder de insertie of de insertie kan onjuist zijn (bijvoorbeeld achteruit). Voordat onderzoekers het recombinant DNA voor verder onderzoek kunnen gebruiken, moeten ze controleren op fouten. Nucleotidensequencing kan helpen bij het identificeren van bacteriekolonies die plasmiden dragen met de juiste insert.



<h4>Wetenschappers gebruiken recombinant DNA om genen en eiwitten te bestuderen</h4>



Recombinant-DNA-technologie is vooral voordelig wanneer een wetenschapper veel kopie&#xEB;n nodig heeft van een gen van interesse of een eiwitproduct. Het onderzoek van een wetenschapper kan echter een extra niveau van complexiteit vereisen, zoals de detectie of zuivering van het gewenste eiwit. Om dit doel te bereiken kan een onderzoeker een tag of reporter &#x2013;eiwitten die worden gebruikt om een genproduct te identificeren&#x2013; aan het gewenste eiwit hechten om een fusiegen of chimeer gen te cre&#xEB;ren.



<h4>Toepassingen in de geneeskunde en de landbouw</h4>



Wetenschappers gebruikten voor het eerst recombinant-DNA-technologie om humane insuline in bacteri&#xEB;n te produceren, wat resulteerde in een behandeling voor diabetes. Sinds die eerste ontdekking hebben onderzoekers andere recombinante DNA&#x27;s gegenereerd voor therapeutisch gebruik. Recombinante bacteri&#xEB;n maken menselijk groeihormoon &#x2013; een eiwit dat nodig is voor normale groei en ontwikkeling &#x2013; om pati&#xEB;nten met groeihormoondefici&#xEB;ntie te behandelen. Recombinante zoogdiercellen, afkomstig van mensen en hamsters, produceren factor VIII &#x2013; een eiwit dat nodig is voor een normale bloedstolling &#x2013; om pati&#xEB;nten met hemofilie te behandelen. Het is duidelijk dat recombinant-DNA-technologie een krachtig hulpmiddel is voor de grootschalige productie van essenti&#xEB;le eiwitten.

De vooruitgang in de landbouw op het gebied van recombinant-DNA-technologie heeft ook invloed op het menselijk welzijn. Zo leden ma&#xEF;sboeren bijvoorbeeld aanzienlijke gewasschade als gevolg van de plaag van de Europese ma&#xEF;sboorder. Als reactie hierop isoleerden wetenschappers genen uit een in de bodem levende bacterie &#x2013; Bacillus thuringiensis (Bt) &#x2013; om genetisch gemodificeerde, ongediertebestendige ma&#xEF;s te cre&#xEB;ren. Bacillus thuringiensis produceert van nature eiwitten die giftig zijn voor bepaalde insecten, maar niet voor mensen, planten of andere dieren. De introductie van pestresistente Bt-ma&#xEF;s verbeterde de gewasopbrengsten en verminderde het gebruik van chemische pesticiden. Dergelijke landbouwtoepassingen verbeteren de kwaliteit en kwantiteit van de mondiale voedselvoorziening.

Recombinant DNA

Overview

Scientists create recombinant DNA by combining DNA from different sources&#x2014;often, other species&#x2014;in the laboratory. DNA cloning allows ...

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recombinant dna in cloning and gmos

Recombinant DNA is made by joining DNA from different sources in the lab. Scientists often use it to clone genes and study them in cells that are easy to handle, such as bacteria. Organisms that carry recombinant DNA are called genetically modified organisms, or GMOs. This technology gives scientists new tools for research, medicine, and agriculture.

To build recombinant DNA, researchers place a gene of interest into a vector, which is a DNA carrier that moves foreign DNA into a host cell. The most common vectors are plasmids, small circular DNA molecules that copy themselves separately from the cell’s chromosomal DNA. Scientists cut both the donor DNA and the vector at specific restriction sites using restriction enzymes. Then DNA ligase seals the sugar-phosphate backbone where the insert joins the plasmid.

The final product is a recombinant DNA molecule with a vector carrying an inserted piece of donor DNA. Scientists can put this hybrid DNA into a host organism, usually bacteria or yeast. The host then replicates the DNA quickly, making many copies of the gene of interest. In some cases, the host’s machinery also transcribes and translates the gene into a protein, such as human insulin.

Creating recombinant DNA is not always perfect. The vector may close without taking in the insert, or the insert may end up in the wrong direction. Before moving on, researchers check for these errors. Nucleotide sequencing helps them find bacteria colonies that contain plasmids with the correct insert.

Recombinant DNA is useful when scientists need many copies of a gene or a protein product. Some projects need extra steps, such as making the protein easier to detect or purify. For that reason, researchers may attach a tag or reporter protein to create a fusion gene, also called a chimeric gene.

This technology has major uses in medicine and agriculture. Scientists first used recombinant DNA to make human insulin in bacteria for diabetes treatment. They also use recombinant bacteria to produce human growth hormone for growth hormone deficiency. Recombinant mammalian cells, from humans and hamsters, make Factor VIII to treat hemophilia. In agriculture, scientists used genes from Bacillus thuringiensis, or Bt, to create pest-resistant corn. Bt proteins are toxic to certain insects, not humans, plants, or other animals, so Bt corn can improve crop yields and reduce chemical pesticide use.

Recombinant DNA in Cloning and GMOs

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key terms and concepts

See how scientists build recombinant DNA with plasmids, restriction enzymes, and ligase, then use it to make proteins and GMOs in medicine and farming.

Recombinant DNA in Cloning and GMOs | JoVE K12

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Recombinant DNA in Cloning and GMOs

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