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Recombinant DNA
Recombinant DNA (rDNA) is an artificial form of DNA intentionally produced with molecular biology techniques that allow disparate sequences of DNA to be joined together. The first use of the rDNA technique dates back to 1973, when Stanley N. Cohen and his colleagues published an article detailing a technique for isolating and amplifying genes or DNA segments and inserting them into bacteria. The discovery that genes isolated from one organism that are transferred into another may be functional has opened the door to further important research in biology, not the least of which is the study of genes and their function in the biological processes of living organisms. Gene transfer techniques also provide the pharmaceutical and agrifood industries with important applications, already a part of daily life. The issues surrounding the modifications introduced in various classes of living organisms from the use of rDNA technologies offer an opportunity for analyzing the involvement of scientific progress in everyday life, the relationship between science and the general public, and the crucial role of communication where science meets society.
Where the genetic characteristics of plants, animals, and microorganisms have been modified artificially, they are defined as genetically modified organisms, sometimes referred to as GMOs. Transgenics is another commonly used term for this technology, while genetic transformation indicates the process of producing such life-forms. The transferred gene is called “foreign” or exogenous and is distinguished from endogenous genes, which are naturally present in the host genome. Recombinant molecules of DNA may be produced from different sources of DNA fragments—whether isolated from different organisms or synthetically generated—that would not normally occur together. For this reason, rDNA is also sometimes referred to as chimeric DNA, after the chimera, a mythological creature whose body is composed of parts of different animals.
Technical Aspects
The production of rDNA is done by cutting DNA sequences into pieces using what are known as restriction enzymes. These are natural scissors of bacterial origin, able to cut DNA at precise points of the molecular strand (restriction sites), which are specific to each enzyme. The resulting genome fragments can be joined again to make new compositions via a reaction named recombination, which involves enzymes called recombinases that are naturally present in microorganisms.
The discovery of restriction enzymes was an important breakthrough in biology and in genetic engineering in particular. The scientists who identified them, Werner Arber, Daniel Nathans, and Hamilton Smith, were awarded the Nobel Prize for Medicine in 1978. Since 1970, when the first restriction enzyme was discovered by this research group, over 3,000 restriction enzymes have been studied and more than 600 of them are now available for routine use in laboratories carrying out DNA modification and manipulation.
Specific sequences are attached to the exogenous gene that are able to switch it on (promoter) and off (terminator) and to allow it to function in the host cell (ribosome binding); after it has been transferred into the new organism, the foreign gene is then capable of coding for a specific recom-binant protein.
Applications
In the last decade, following several genome-sequencing projects aimed at understanding genome structures and identifying the genes of various microorganisms, plants, and animals, including humans, a huge amount of data has been collected. The next step is to apply this basic knowledge to understanding gene functions and interactions (functional genomics) using—in addition to laboratory and computer-based analyses—rDNA techniques, which offer the possibility of transferring DNA sequences into living organisms for more extensive and reliable analysis. Functional genomics based on gene transfer is becoming one of the most promising aspects of genetic research today, especially in microorganisms and plants.
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