Recombinant DNA Technology & Cloning
RECOMBINANT DNA TECHNOLOGY
Recombinant DNA Technology refers to the process by which DNA molecules of two different species are joined together and then inserted into a host for the production of new genetic combinations which are valuable to science, medicine, agriculture and industry.
Steps involved in this process are:
The genetic material in living organisms is generally nucleic acids. Whereas in most of the cases it is DNA and sometimes it is RNA. The first step in the process of RDT is to isolate the desired DNA in free form i.e. free from other macromolecules. So as DNA is bounded in the cell membrane along with several other molecules, enzymes are required to break all those barriers for its isolation which can be:
After the removal of the other unnecessary particles, addition of ethanol allows DNA to precipitate out as fine threads.
The restriction enzymes act as molecular scissors that cut DNA at specific sites. Agarose Gel Electrophoresis is performed in this process. Here, DNA is run on agarose gel and current is passed. In result, negatively charged DNA moves to positive electrode and settles according to size. This allows the separation and cut of digested DNA fragments.
In PCR, multiple copies of DNA is made using enzyme DNA Polymerase. It helps to amplify few copies of DNA into millions of copies.
Purified DNA and vector of interest are cut using restriction enzymes. This gives the cut DNA and vector fragments. The process of joining these is Ligation with the enzyme DNA ligase. The resulting DNA is recombinant DNA.
Transformation is the process by which recombinant DNA is introduced into a host. Also the bacterial cells should be treated to make them competent to accept new DNA.
New DNA multiplies inside the host and is expressed as protein. This is now a recombinant protein.
Before the protein is released it is subjected to downstream processing that includes:
DNA Cloning is an important process in Recombinant DNA Technology that has provided the scientists with the ability to produce many copies of a single fragmented DNA. This is done by inserting a small fragment of DNA into a DNA molecule which is allowed to replicate in a growing living cell such as a bacterium. The small replicating molecule is a DNA vector or carrier. The most commonly used vectors are plasmids, viruses and yeast cells.
Steps involved in this process:
Restriction Enzymes are used for cutting DNA fragments.
Digestion is done
DNA Ligase is used to combine fragments that allow them to link and make a recombinant plasmid containing the gene.
During transformation, specially prepared bacterial cells are given a shock that allows them to take up the foreign DNA.
After the bacteria get DNA they form colonies.
After the bacteria form colonies they are given a chemical signal that instructs them to produce target proteins. Once it forms, bacterial cells can be split open to release it. The proteins are then purified and can be used in various experiments.
Recombinant DNA Technology is useful in Industrial purposes in terms of commercial importance, improvement of fermentation processes and production of proteins from wastes.
Most of the cons of Recombinant DNA Technology are ethical in nature. According to some of the organizations, it is seen to have felt that this technology goes against the laws of natures or against the religious beliefs. Some worries are also been seen that if companies can allow scientist with patent to buy or sell genetic materials then it could become an expensive commodity. Problems with the safety of modified foods and medicines also occur.
What is cloning? There are 3 main types of cloning (Genetics Generation, n.d), Reproductive, Gene and therapeutic cloning. Gene cloning is when a copy of DNA is inserted into a vector which is then able to be copied by the host, therapeutic cloning is when stem cells are cloned to treat disease and for research and Reproductive cloning which is the creation of an animal identical to the donor and the one most referred to when talking about cloning which...
Introduction In the advanced world, consistently there has been a remarkable social tool for comprehending and assessing scientific, ethical, moral, and social consequences of new innovations. Cloning is one such new technology which is a major development in the field of Genetics, such as Human Genome Project for making identical copies of an organism. Cloning is the term broadly meaning natural or artificial development of two or more genetically identical cells or organisms. Scientists use the term cloning with variable...
Is Human Cloning Worth the Expense of Further Research? Introduction What is Human Cloning? Human cloning, refers to the process of creating a human being that is genetically identical to a pre-existing person through the use of their cells. (Science Daily, 2019), (Center for Genetics and Society, 2019). Despite many scientist claiming to have done so, there is no verified experiment that has actually cloned another human being. The process of cloning called somatic cell nuclear transfer (SCNT) was used...
In the wake of fast scientific and technological advancements witnessed in the 21st Century, much has been debated concerning human cloning and the use of technology. Most of the discussions have centered on different religions that provide different views regarding cloning and the use of Assisted Reproductive Technology (ART). Human cloning has raised different controversial sights in Christianity, Buddhism, Protestant Churches, Islam, and many others. The religions that oppose human cloning are of the view that it is a practice...
Gene cloning is a biotechnology in which a section of DNA is isolated and extracted before being cloned using bacterial plasmids. To execute this process, a restriction enzyme isolates a specific gene from a strand of DNA and the plasmid, and then cuts the gene creating complementary sticky or blunt ends. These are joined to form recombinant DNA, which is then inserted into bacteria via heat shock to be cloned. Gene cloning is used for many reasons such as to...
Humans have been breeding dogs for at least an estimated 14,000 years, it is said to believe that the evolution of the domestic dog evolved from the wild grey wolf (Canis Lupus.) As humans began domesticating dogs, they had begun to favour specific traits such as dogs with particular physiques and temperaments, hunting skills, intelligence, behavioural traits and even the simple companionship between man and dog.By selecting the most favourable dogs to breed, humans were unconsciously using artificial selection to...
It basically involves a method for developing a replica of tissue, organ or cell which are inherently similar to each other.it occur in nature- for instance, when some cell replicated itself asexually without chromosomal mutation and linkage. In prokaryotes, bacteria can produce a genetically alike copy of itself with help of binary fission. On other hand, in eukaryotes like in humans all cells such as skin cells, digestive track lining cells go through mitosis thus produce clones, except from the...
As scientists try to understand the existence of DNA and genetics, they soon discover the concept of cloning in biotechnology. Cloning is a process that creates exact copies of DNA on different molecular levels. For example, there is molecular cloning where copies of DNA fragments are made. There is also cell cloning, as well as organismal cloning. Moreover, there are three different types of cloning that are currently known in biotechnology. First, there is recombinant DNA technology which is also...
‘To what extent does animal cloning breach morality and under what conditions can it be justified?’ Does potential warrant use of unsatisfactory methods? The future of animal cloning, a process of generating a genetically identical copy of a cell or an organism, depends on the answer to this poignant question. Since its inception in 1996 with the cloning of Dolly the sheep from an adult somatic cell, clone production has been a very controversial topic and the allowance of this...
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