Most important to mention are the new approaches utilized for waste treatments and bioremediation Environment protection means the conservation of resources and hence to limit the degradation of environment. The advances in genetic engineering are of immense help for humanity but if mishandled, their prospects are quite frightening. Visualization of a specific DNA (or RNA or protein) fragment out of many molecules requires a technique called blot transfer. In simple words it is the introduction of recombinant DNA molecule into a host cell which is then multiplied to produce the clones of rec DNA. (vii) Host bacterium divides to give multiple copies of the recombinant DNA which may be preserved in a DNA library which is also called genomic DNA library. First success in the approach of artificial gene synthesis was achieved by Dr. Har Govind Khorana and his co-workers in 1970 when they synthesized the artificial gene for a t-RNA in vitro which had potential for functioning within a living cell. In this article we will discuss about the steps and applications of Recombinant DNA Technology. of the nitrogen bases can be arranged in an infinite ways, and their structure is known as . 4. 3. Roundup Ready soya been developed by. Why does plant cell possess large sized vacuole? Recombinant DNA Technology. It is actually a gene manipulation where one DNA molecule is attached to another. Some improved traits generally present in transgenic plants are one or more from the following (Table 2): (iii) Resistance against abiotic stresses like: (iv) Modified quality of starch, edible-oils, proteins obtained from crop plants. It is true that biotechnology and genetic engineering have immense applications in almost all the areas related to the betterment of humanity. Hazardous toxins can be produced by genetic engineering of several organism like botulinum toxin, neurotoxins, aflatoxins etc.. can be used as biological weapons. 7. In cases where the samples of blood stain, semen etc. In case of plants, such probes are not present, therefore, usually RFLPs (Restriction Fragment Length Polymorphism) or simple sequence repeats like (CT) or (AC)n etc. A genetically engineered soybean manufactured by a British company was banned as it caused allergies in some persons on consumption. Following the migration of the molecules, the gel is treated with selective stains to show the location of separated molecules in the form of bands. Recombinant DNA (rDNA) molecules are DNA molecules formed by laboratory methods of genetic recombination (such as molecular cloning) to bring together genetic material from multiple sources, creating sequences that would not otherwise be found in the genome. Applications of Recombinant DNA Technology. The development of rec DNA technology specially the use of restriction enzymes has provided an extra boost to Human Genome Project. These may be present in bacterial clones with recombination DNA or by obtaining complementary DNA (c-DNA) by obtaining a DNA from an RNA template by using the enzyme reverse transcriptase. Major limitation of PCR is that due to its extreme sensitivity it may produce erroneous results caused by several inhibitors or contaminating DNA segments present in the sample DNA preparation. Such cuts are termed as staggered cuts and this results into the generation of protruding ends i.e., one strand of the double helix extends a few bases beyond the other strand. Human Genome project (HGP) is an ambitious plan which is administered by National Institute of Health and Department of Energy, U.S.A. 7. Transgenic research is being carried over in India also with a view to strengthen the plant biotechnology in country. There should not be any interfering element against the replication of rec DNA in the host cells. Monoclonal antibodies are useful tools for disease diagnosis. This can be done by following any of the gene delivery systems available like AMGT (Agrobacterium mediated gene transfer), using plant viruses as vectors or by direct gene delivery system i.e., electroporation, microinjection, particle-gun method, etc. It can be used to elucidate molecular events in the biological process of cellular differentiation and ageing. Other DNA are techniques available for breaking a DNA molecule into shorter fragments. For sequencing, the reaction mixture is taken in four separate test tubes. 4. Such types of restriction endonucleases cleave the DNA about 1000 bp away from the 5′ end of the sequence ‘TCA’ located within the recognition site. (iv) rec DNA molecule is introduced into a suitable host: Suitable host cells are selected and the rec DNA molecule so formed [in step (iii)] is introduced into these host cells. In plant molecular farming, first of all the plants to be used as the ‘expression system’ are chosen. Also, the transformed host cells are multiplied to obtain sufficient number of copies. c. A heat stable DNA polymerase e.g. Transgenic animals also contribute for studying the gene functions in different animal species. A foreign gene associated with the production of desired biomolecule is then integrated into plants’ genome. Gene drug preparation and Gene therapy approaches involve the introduction of genes into the target cells. (With Methods)| Industrial Microbiology, How is Cheese Made Step by Step: Principles, Production and Process, Enzyme Production and Purification: Extraction & Separation Methods | Industrial Microbiology, Fermentation of Olives: Process, Control, Problems, Abnormalities and Developments, The best answers are voted up and rise to the top. Yeast: Origin, Reproduction, Life Cycle and Growth Requirements | Industrial Microbiology, How is Bread Made Step by Step? However, often this term i.e., gene cloning is used as a synonym to rec DNA technology or genetic engineering. Steps in Recombinant DNA Technology: Basic steps involved in rec DNA technology (or genetic engineering) … U.S. scientists reported the production of a super- AIDS virus which was formed when the ordinary AIDS virus was combined with the mice virus. One such enzyme is DNA polymerase ! prevention of a number of diseases like AIDS, cholera, etc. The desired DNA segment which is to be cloned is called as DNA insert or foreign DNA or target DNA. complementary DNA from the mRNA of that gene. 8. The fermenting microbes which are utilized for biogas production are improved at the genetic level for achieving better result. PvuII, Haelll, Alul are the examples of restriction endonucleases producing blunt ends. 6. Genetic engineering plays an essentially important role in a beneficial and large scale production of biofuels like biogas. One or more deoxynbonucleotides (dATP, dGTP, dl IP, dCTP) are added onto the 3′-end of the blunt-ended fragments. To identify virus & bacteria in infectious diseases. As a result, different sizes of newly synthesized DNA strands are obtained in each test tube which are terminated by a particular ddNTP. It is another important natural tool which geneticists use in rec DNA technology. Usually agarose gel is used for separation of large segments of DNA while the polyacrylamide gel is used for the separation of small DNA fragments which are only a few base pairs long. A number of institutes like Central Potato Research Institute (CPRI), Shimla; Indian Agricultural Research Institute (IARI), New Delhi; Central Rice Research Institute (CRRI), Cuttack; Delhi University, Punjab University, Ludhiana, etc. 1): iii. High lauric rapeseed is an approved genetically engineered plant which produced rapeseeds rich in laurate (fatty-acid) which is useful in making soaps, detergents and shampoos. Rhizobium melilottii is a successful example in this case. Advancements in biotechnology has also aided the terrorists to produce potent biological warfare agents. This enables better characterization of DNA which would not be possible otherwise. Choose the best answer from the four options given. The methods commonly used for DNA sequencing are: i. Enzymatic method or Sanger’s Dideoxy method. They are the intermediate enzymes between Type-I and Type-II restriction endonuclease. The detection of integration of foreign genes into the plant genome requires the use of marker gene that either allow the transformed cells to be selected (selectable marker gene) or that encodes an activity which can be assayed or scored (scorable marker gene). These methods differ in their strategies for protecting the hydroxyl group of the phosphate residues. Such techniques are faster and more reliable. Some important antibiotics are tetracyclin, penicillin, streptomycin, novobiocin, bacitracin, etc. From the above discussion it is clear that the field of biotechnology especially the genetic engineering is a double edged sword. 5b). These are also applied for studying the regulation of gene expressions under different conditions of factors like light and temperature (Table 2). But there are certain risks/harms related to it also It has been found that biotechnological processes may also have adverse effects in several areas. Recombinant DNA Research. It can be used to elucidate molecular events in the biological process of cellular differentiation and ageing.