Recombinant DNA technology is a biotechnology approach that has multidisciplinary applications and the potential to deal with important aspects of life, from health issues (e.g. by the means of recombinant antibodies) to food resources, and resistance to divergent adverse environmental effects. As genetic diseases are becoming more prevalent and agricultural areas are being reduced, the technology is gaining in importance in fields such as biochemistry and microbiology.
In this article, we will show you the steps in recombinant DNA production, methods and examples as well as outlooks in recombinant DNA technology.
Recombinant DNA technology is, as the name already suggests, a means to generate recombinant DNA. This process consists of the manipulation as well as the isolation of specific DNA segments from different species. DNA is joined by attaching segments of the molecule to the DNA of a virus or bacterial plasmid (to be found in bacterial cells), and subsequently inserted into a host cell, often a bacterial or yeast cell.
Plasmids, artificial chromosomes or bacteriophages are widely used as vectors in molecular cloning, serving to drive the replication of recombinant DNA sequences within host organisms – living cells.
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There are several pros and cons of recombinant DNA. The disadvantages mainly relate to ethical or religious concerns in the commercialization of products containing recombinant DNA. But genetic engineering offers great opportunities in the research and production of innovative medical or therapeutic products.
While the structure of DNA was first determined in 1953, it wasn’t until the early 1970 years that the first recombinant DNA molecules were produced by the means of restriction enzymes. Paul Berg (Stanford) succeeded in proving the possibility to splice and to recombine genetic material in 1971.
Consequently, Stanley Cohen (Stanford) and Herbert Boyer (UC San Francisco) were able to cut and fuse specifically located DNA strands – a subsequently submitted patent for recombinant antibody technology was approved in 1980.1
DNA replication can be applied in a range of fields, from medical to biochemical. In the following, we will give you some examples.
The first commercial healthcare product derived from rDNA was human insulin. Today, it is successfully applied to make new antibodies, vaccines (e. g. for Hepatitis B) and different protein production systems, for instance for insulin and human growth hormone.2
Here are 5 examples of rDNA technology in the health sector:
Recombinant DNA technology is used to genetically modify plants in order to improve adaptability as well as resistance to harmful agents and to enhance product yield.2
Examples of rDNA technology in agriculture include:
Recombinant DNA technology enables the manufacturing of novel enzymes that are suitable to prolong shelf life and kill foodborne pathogens.2
Here are 3 examples of recombinant DNA technology in the food industry:
Read more: Application of rDNA technology: What is recombinant DNA used for?
Recombinant DNA technology describes a process of genetic engineering that uses enzymes and various laboratory techniques to isolate and manipulate genetic material in various steps. This is possible because DNA molecules from all organisms share the same chemical structure, and differ only in the nucleotide sequence. There are different ways to carry out manipulation in an organism’s genome, all with the aim to improve certain characteristics.
Recombinant DNA differs from genetic recombination in that it results from artificial methods in the test tube, while the latter is a normal biological process that results in the remixing of existing DNA sequences in essentially all organisms.
There are three different methods of producing recombinant DNA, namely transformation, non-bacterial transformation, and phage.
Transformation and non-bacterial transformation are similar processes, with the only difference being the use of bacteria such as E. Coli for the host. While the transformation process uses Escherichia coli to act as a biological framework, the non-bacterial method does not use any bacteria – as is evident in the name. Initially, one needs to choose which DNA segment to insert into the vector. The second step is to cut that piece of DNA with a restriction enzyme (restriction endonuclease) and then ligate the DNA insert into the vector with DNA Ligase. The vector is inserted into a host cell, which has to be specifically prepared to take up the foreign DNA.
Phage introduction is the process of transfection, which is equivalent to transformation, but in this process, a phage is used instead of bacteria. The in-vitro process uses lambda or MI3 phages to produce phage plaques containing recombinants. By the means of different selection methods, the generated recombinant DNA can then be differentiated from non-recombinant DNA.
Recombinant DNA is composed of sequences that are derived from different sources. The process to achieve this involves the following steps:
The last step can be achieved by an immunological method or nucleic acid hybridization, blue-white screening or insertional inactivation.
The DNA sequences used in the construction of recombinant DNA molecules can originate from any eukaryotic species, be it human, bacterial, fungal or mammalian. In addition, DNA sequences that do not occur naturally may be created by chemical synthesis. The different molecular biology tools used in rDNA technology include DNA isolation and analysis, molecular cloning, quantification of gene expression, determination of gene copy number, transformation of the appropriate host for replication starting at a selectable marker, or transfer into crop plants and analyses of transgenic plants.
Respectively, some of the most important instruments needed are enzymes, gene cloning vectors, polymerase chain reaction (PCR) and host organisms. Eligible host organisms for recombinant antibody expression include bacterial and yeast cells, but also insect or mammalian cells, such as HEK293 cells or CHO cells.
When considering the pros and cons of recombinant DNA, safety is a frequently discussed aspect. Generally speaking, recombinant DNA molecules and recombinant proteins are not regarded as dangerous. However, concerns remain about some organisms that express recombinant DNA, particularly when they leave the laboratory and are introduced into the environment or food chain. Such potential safety issues include antibiotic resistance and adverse immune reactions. Outside the health sector concerns include the potential of gene pollution of the environment but also health effects of foods from GMOs.
However, to ensure the greatest degree of safety possible, all rDNA work needs to be compliant with standards and guidelines set out by the FDA and other regulatory institutions.
Apart from safety issues, there is the ethical issue of genetically modified organisms, human genome editing as well as around genetic information in general.
Today, recombinant DNA technology plays a vital role in improving health conditions by developing new vaccines and gene therapy products but also in dealing with several plant disorders, especially viral and fungal resistance.
With the rising prevalence of chronic diseases being one of the main driving factors, the size of the global recombinant DNA technology market is forecast to reach USD 223 billion by 2028, with an annual growth rate of 7.7% during the forecast period.
Recombinant DNA technology is a laboratory technique that involves manipulating DNA fragments from different organisms to create new genetic combinations that would not be found in nature. It has various applications, including the production of therapeutic proteins and genetically modified organisms. Read more: What is recombinant DNA?
The steps in recombinant DNA technology include: isolating DNA from the donor and host organisms, cutting the DNA using restriction enzymes, joining the fragments with DNA ligase, introducing the recombinant DNA into the host organism, and selecting and screening transformed cells.
Examples of recombinant DNA technology include the production of human insulin, genetically modified crops, gene therapy, production of vaccines (such as the hepatitis B vaccine), and creation of transgenic animals. Read more: 5 examples of recombinant DNA technology
No, recombinant DNA and GMO (Genetically Modified Organism) are not the same. Recombinant DNA refers to DNA molecules formed through combining genetic material from different sources, while GMOs are organisms whose genetic material has been altered using recombinant DNA technology.
Transgenic organisms are organisms whose genetic material has been modified by the introduction of genes from another species using recombinant DNA technology. These inserted genes can come from plants, animals, bacteria, or other sources. The purpose of creating transgenic organisms is often to confer specific desirable traits, such as increased resistance to pests, improved nutritional content, or enhanced growth rates. Transgenic organisms are commonly referred to as genetically modified organisms (GMOs) and have applications in agriculture, medicine, and research.
Recombinant DNA technology is performed according to specific processes, usually involving the following steps: