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Recombinant DNA (rDNA)

1973
  • Stanley Norman Cohen
  • Herbert Boyer
Biologo molecolare che conduce la tecnologia del DNA ricombinante in laboratorio.

(Immagine generata a solo scopo illustrativo)

Recombinant DNA (rDNA) technology involves joining together DNA molecules from two different species. The recombinant DNA is inserted into a host organism to produce new genetic combinations. This is achieved using restriction enzymes to cut DNA at specific sites and DNA ligase to join the fragments, often incorporating the desired gene into a plasmid vector for cloning.

Recombinant DNA technology, also known as genetic engineering, is the process of creating artificial DNA by combining genetic material from different sources. This technology fundamentally changed biology and medicine by allowing scientists to directly manipulate the genetic code of organisms. The core procedure involves several key steps. First, a gene of interest is identified and isolated from a source organism’s DNA. This is often done using restriction enzymes, which are proteins that act like molecular scissors, cutting DNA at specific recognition sequences. Second, a vector, which is a DNA molecule used to carry the foreign genetic material into another cell, is chosen. Bacterial plasmids—small, circular DNA molecules separate from the bacterial chromosome—are the most common vectors. The same restriction enzyme used to cut out the gene is used to cut open the plasmid vector. This creates compatible ‘sticky ends’ on both the gene and the plasmid. Third, the isolated gene is inserted into the plasmid. The sticky ends of the gene anneal with the complementary sticky ends of the plasmid, and the enzyme DNA ligase is added to permanently join them by forming phosphodiester bonds. The resulting molecule is a recombinant plasmid containing the new gene. Finally, this recombinant vector is introduced into a host organism, typically a bacterium like *E. coli*, through a process called transformation. As the host cells multiply, they replicate the recombinant plasmid along with their own DNA, creating many copies of the inserted gene. The host cells can also transcribe and translate the foreign gene to produce the desired protein, such as human insulin produced in bacteria.

UNESCO Nomenclature: 2406
- Biologia molecolare

Tipo

Processo chimico

Interruzione

Rivoluzionario

Utilizzo

Uso diffuso

Precursori

  • discovery of DNA as genetic material
  • elucidation of the DNA double helix structure
  • discovery of plasmids in bacteria
  • discovery and characterization of restriction enzymes by Werner Arber, Daniel Nathans, and Hamilton Smith
  • discovery of DNA ligase

Applicazioni

  • production of synthetic human insulin for diabetics
  • creation of genetically modified organisms (GMOs)
  • production of vaccines (e.g., hepatitis B vaccine)
  • gene therapy
  • production of clotting factors for hemophilia

Brevetti:

  • US4237224

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Argomenti correlati: DNA ricombinante, ingegneria genetica, Stanley Cohen, Herbert Boyer, plasmide, vettore, enzima di restrizione, DNA ligasi, clonazione genica, OGM.

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Recombinant DNA (rDNA)

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