Recombinant DNA Technology
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Abstract: Genethics actually is a 1990 book by David Suzuki and Peter Knudtson, published by Harvard University Press about The Clash between the New Genetics and Human Values. At present the technologies used are vast say we have recombinant DNA technology , gel electrophoresis , gene mapping, gene mutation, DNA sequencing , etc . These technologies have vast applications in fields of medicine, genetics etc. The future of genetic technology is really very critical for this already suffering planet EARTH.If this technology falls in wrong hands then future certainly looks bleak.
Genetic engineering alters the genetic makeup of an organism using techniques that introduce heritable material prepared outside the organism either directly into the host or into a cell that is then fused or hybridised with the host
Genethics speaks about the definition basically then it tells us the history of genetic engineering, that had triggered the minds of many people who took interest in the experiments conducted in
the past and gave it another try and became successful.
Technologies used are recombinant DNA technology , cloning , gene mapping, gene mutation, DNA sequencing.
Karthik V Sushil Baranwal
SIES GST, Nerul SIES GST, Nerul
RECOMBINANT DNA TECHNOLOGY
Recombinant DNA (rDNA) is a form of artificial DNA that is created by combining two or more sequences that would not normally occur together. In terms of genetic modification, it is created through the introduction of relevant DNA into an existing organismal DNA, such as the plasmids of bacteria, to code for or alter different traits for a specific purpose, such as antibiotic resistance. It differs from genetic recombination in that it does not occur through natural processes within the cell, but is engineered. A recombinant protein is a protein that is derived from recombinant DNA.
The recombinant DNA technique was first proposed by Peter Lobban, a graduate student, with A. Dale Kaiser at the Stanford University Department of Biochemistry "Biochemical Method for Inserting New Genetic Information into DNA of Simian Virus 40: Circular SV40 DNA Molecules Containing Lambda Phage Genes and the Galactose Operon of Escherichia coli".
Cloning in biology is the process of producing similar populations of genetically identical individuals that occurs in nature when organisms such as bacteria, insects or plants reproduce asexually. Cloning in biotechnology refers to processes used to create copies of DNA fragments (molecular cloning), cells (cell cloning), or organisms. The term also refers to the production of multiple copies of a product such as digital media or software.
Molecular cloning refers to the process of making multiple molecules. Cloning is commonly used to amplify DNA fragments containing whole genes, but it can also be used to amplify. any DNA sequence such as promoters, non-coding sequences and randomly fragmented DNA.
Cloning of any DNA fragment essentially involves four steps
fragmentation - breaking apart a strand of DNA
ligation - gluing together pieces of DNA in a desired sequence
transfection - inserting the newly formed pieces of DNA into cells
screening/selection - selecting out the cells that were successfully transfected with the new DNA .
A . Horizontal gene transfer (HGT), also Lateral gene transfer (LGT), is any process in which an organism incorporates genetic material from another organism without being the offspring of that organism. By contrast, vertical transfer occurs when an organism receives genetic material from its ancestor, e.g., its parent or a species from which it has evolved.
Most thinking in genetics has focused upon vertical transfer, but there is a B. Particle Gun Method - In this method, 1-2µm tungsten or gold particles, coated with the DNA to be used for transformation, are accelerated to velocities, which enable their entry into plant cells/nuclei. Particle aceleration is achieved by using a device, which varies considerably in design and function.
The most successful devices accelerate particles in one of the two ways:
(1) by using pressurised helium gas or
(2) by the electrostatic energy released by a droplet of water exposed to a high voltage.
STEM CELL TECHNIQUE
stem cell transplantation (SCT) is the transplantation of Pluripotential stem cell or blood, often derived from bone marrow, umbilical cord blood or hemopoietic stem cells derived from a placenta. Stem cell transplantation is a medical procedure in the fields of hematology and oncology, most often performed for people with diseases of the blood, bone marrow, or certain cancer.
With the availability of the stem cell growth factors GM-CSF and G-CSF, most hematopoietic stem cell transplantation procedures are now performed using stem cells collected from the peripheral blood such as cord blood or placenta derived stem cells, rather than from the bone marrow. Collecting peripheral blood stem cells provides a bigger graft, does not require that the donor be subjected to general anesthesia to collect the graft, results in a shorter time to engraftment, and may provide for a lower long-term relapse rate.
stem cell transplantation remains a risky procedure with many possible complications; it has traditionally been reserved for patients with life-threatening diseases. While occasionally used experimentally in nonmalignant and nonhematologic indications such as severe disabling auto-immune disease and cardiovascular disease, the risk of fatal complications appears too high to gain wider acceptance.
DNA profiling (also called DNA testing, DNA typing, or genetic fingerprinting) is a technique employed by forensic scientists to assist in the identification of individuals on the basis of their respective DNA profiles. DNA profiles are encrypted sets of numbers that reflect a person's DNA makeup, which can also be used as the person's identifier. DNA profiling should not be confused with full genome sequencing. It is used in, for example, parental testing and rape investigation.
The process begins with a sample of an individual's DNA (typically called a "reference sample"). The most desirable method of collecting a reference sample is the use of a buccal swab, as this reduces the possibility of contamination. When this is not available (e.g. because a court order may be needed and not obtainable) other methods may need to be used to collect a sample of blood, saliva, semen, or other appropriate fluid or tissue from personal items (e.g. toothbrush, razor, etc.) or from stored samples (e.g. banked sperm or biopsy tissue). Samples obtained from blood relatives (biological relative) can provide an indication of an individual's profile, as could human remains which had been previously profiled.
The DNA profile is then compared against another sample to determine whether there is a genetic match.
The term DNA sequencing refers to sequencing methods for determining the order of the nucleotide bases-adenine, guanine, cytosine, and thymine-in a molecule of DNA.
Knowledge of DNA sequences has become indispensable for basic biological research, other research branches utilizing DNA sequencing, and in numerous applied fields such as diagnostic, biotechnology, forensic biology and biological systematics. The advent of DNA sequencing has significantly accelerated biological research and discovery. The rapid speed of sequencing attained with modern DNA sequencing technology has been instrumental in the sequencing of the human genome, in the Human Genome Project. Related projects, often by scientific collaboration across continents, have generated the complete DNA sequences of many animal, plant, and microbial genomes.
The classical chain-termination method requires a single-stranded DNA template, a DNA primer, a DNA polymerase, radioactively or fluorescently labeled nucleotides, and modified nucleotides that terminate DNA strand elongation. The DNA sample is divided into four separate sequencing reactions, containing all four of the standard deoxynucleotides (dATP, dGTP, dCTP and dTTP) and the DNA polymerase.
Tadpole: (1952) Many scientists questioned whether cloning had actually occurred and unpublished experiments by other labs were not able to reproduce the reported results.
Carp: (1963) In China, embryologist Tong Dizhou produced the world's first cloned fish by inserting the DNA from a cell of a male carp into an egg from a female carp. He published the findings in a Chinese science journal.
Mice: (1986) A mouse was the first successfully cloned mammal. Soviet scientists Chaylakhyan, Veprencev, Sviridova, and Nikitin had the mouse "Masha" cloned. Research was published in the magazine "Biofizika" .
Sheep: (1996) From early embryonic cells by Steen Willadsen. Megan and Morag cloned from differentiated embryonic cells in June 1995 and Dolly the sheep from a somatic cell in 1997.
The first human hybrid human clone was created in November 1998, by American Cell Technologies. It was created from a man's leg cell, and a cow's egg whose DNA was removed. It was destroyed after 12 days. Since a normal embryo implants at 14 days, Dr Robert Lanza, ACT's director of tissue engineering, told the Daily Mail newspaper that the embryo could not be seen as a person before 14 days. While making an embryo, which may have resulted in a complete human had it been allowed to come to term, according to ACT: "[ACT's] aim was 'therapeutic cloning' not 'reproductive cloning'" .
Transgenic plants have genes inserted into them that are derived from another species. The inserted genes can come from species within the same kingdom (plant to plant) or between kingdoms (bacteria to plant). In many cases the inserted DNA has to be modified slightly in order to correctly and efficiently express in the host organism. Transgenic plants are used to express proteins like the cry toxins from Bacillus thuringiensis, herbicide resistant genes and antigens for vaccinations.
Cisgenic plants are made using genes found within the same species or a closely related one, where conventional plant breeding can occur. Some breeders and scientists argue that cisgenic modification is useful for plants that are difficult to crossbreed by conventional means (such as potatoes), and that plants in the cisgenic category should not require the same level of legal regulation as other genetically modified organisms
Genetic Engineering has caused much controversy in the scientific world as well as amidst the general population. Although the fear of terrorism and cloning is instilled amidst most, there are many positive aspects to this sport which makes it very plausible. There are many medical aspects such as cures for diseases and enhanced food. Genetic Engineering is slowly progressing as laws and policies of different regions slowly begin to accept the many possibilities and dispose of the negative aspects.
The genetically modified foods controversy is a dispute over the relative advantages and disadvantages of genetically modified (GM) food crops and other uses of genetically-modified organisms in food production. The dispute involves biotechnology companies, governmental regulators, non-governmental organizations and scientists. The dispute is most intense in Japan and Europe, where public concern about GM food is higher than in other parts of the world such as the United States. In the United States GM crops are more widely grown and the introduction of these products has been less controversial.
The five key areas of controversy related to genetically engineered food are food safety, the effect on natural ecosystems, gene flow into non GE crops, moral/religious concerns, and corporate control of the food supply.
The Bt Brinjal Controversy
Bt Brinjal is a transgenic brinjal created by inserting a gene from the soil bacterium Bacillus thuringiensis into Brinjal. The insertion of the gene into the Brinjal cell in young cotyledons has been done through an Agro bacterium-mediated vector, along with other genes like promoters, markers etc. This is said to give the Brinjal plant resistance against lepidopteron insects like the Brinjal Fruit and Shoot Borer (Leucinodes orbonalis) and Fruit Borer (Helicoverpa armigera). The transgenic brinjal was developed by Mahyco (Maharashtra Hybrid Seeds Company) in collaboration with the US-based transnational, Monsanto. Release of Bt brinjal into the environment for food, feed and cultivation may present a serious risk for human and animal health; the GM brinjal is unfit for consumption. Bt cotton hybrids require more water than the traditional varieties (31). In a predominantly rain-fed agrarian economy, high water requirements may destroy many GM crops as well as deplete already scarce ground water sources.
The future of genetic engineering is contained in both an awesome yet uneasy picture. There are many moral issues involved with genetic engineering in all areas. Huge numbers of people are both against and supportive of human cloning. Some believe that is unethical and unfair to clone human beings, and yet others desperately want to see it happen. And although many are against it, and even though the United States has banned human cloning, scientists predict that in the future, if not soon, human cloning will occur.
As of current, the main uses for genetic engineering are medicine, industry, and agriculture. But there are many other uses that are in the future for genetic engineering. In agriculture, many more foods are being considered for genetic changes or additions. But the specifics of each food's case can not be presently determined, since not a single gene of an organism would pass on all traits of the organism. Fish genes can be combined with a type of produce and the produce would not take on any physical or otherwise characters of the fish, except for maybe growth or such. There is slight unrest however, in that many people believe the long time effect of genetically produced foods will have a negative side, and so if that is the case, then genetic engineering in agriculture and foods would probably downslide, if not lose all credit. Genetic engineering in industry also has many future uses. But like any new area, there are positives and negatives. So any advance in maybe bio-war products or such, might evoke opposition from society, even if there are benefits. Genetic engineering in medicine is probably the most exciting area. There are current attempts through studying genetics to determine how certain people would react to certain drugs. These studies could eventually lead to a higher level of genetic engineering in medicine, to prepare drugs especially for a person based on their genetic information. Even though distant now, the far future holds the prospect of reaching the eventual goal of being able to determine what disease a human has before the disease strikes. Then if the disease is a complex one, one that relies on the environment around the person, changes can be made to prevent the disease. Also, there are hopes of being able to find more cures and preventions for diseases.
The future is unknown in specifics, yet the dreams and hope of scientists and common society soar far ahead in time. With dedicated research, inspired scientists, and hard work, what might be set for ahead of us, could be brought to us sooner.
A new technology is dawning on our era, a technology that could change the lives of our children and their children to come. Will this medical advancement be pursued or will it be outlawed? The medical advancement is genetic engineering, the duplicating of human cells, and people have strong feelings both for and against it. Many people who have strong feelings for why cloning should be allowed say, " Cloning human cells could one day save your life and the lives of many other people. This research could find cures for cancer, genetic diseases such as cystic fibrosis, and damaged hearts, livers, and brains" . On the other hand, the people who feel otherwise say, "Introducing genes into chromosomes is very much a hit or miss proposition. Scientists might achieve the results they intended once in twenty times, making the procedure far to risky to preform on a human embryo" .
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