Recombinant DNA Technology and Restriction Endonucleases by Juan Carlo Flordeliza https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p SGD #6 en-us 2020-10-26 06:04:32 UTC 2025-04-06 18:32:29 UTC hello@padlet.com https://padlet.net/icons/png/1f9ec.png PRINCIPLES OF RECOMBINANT DNA TECHNOLOGY juancarloflordeliza https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/860581896 1. Isolation of Genetic Material: Isolation of the desired DNA in its pure form i.e. free from other macromolecules.
2. Restriction Enzyme Digestion: incubation of the purified DNA with the selected restriction enzyme, at conditions optimal for that specific enzyme.
3. Amplification Using PCR: Polymerase Chain Reaction or PCR is used to amplify a single copy or a few copies of DNA into thousands to millions of copies
4. Ligation of DNA Molecules: The purified DNA and the vector of interest are cut with the same restriction enzyme 'Ligation". The result­ing DNA molecule is a hybrid of the interest molecule and the vector 'Recombination'
5. Insertion of Recombinant DNA Into Host: The recombinant DNA is introduced into a recipient host cell mostly, a bacterial cell ‘Transformation’
6. Isolation of Recombinant Cells: The transformation process generates a mixed population of transformed and non-transformed host cells. This selection process involves filtering the transformed host cells only. For isolation of recombinant cell from non-recombinant cell, a marker gene of plasmid vector is employed

Source: Recombinant DNA Technology- Steps, Applications, and Limitations (https://microbenotes.com/recombinant-dna-technology-steps-applications-and-limitations/)

]]> 2020-10-26 06:07:32 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/860581896 RECOMBINANT DNA TECHNOLOGY juancarloflordeliza https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/860582599 Recombinant DNA technology comprises altering genetic material outside an organism to obtain enhanced and desired characteristics in living organisms or as their products. This technology involves the insertion of DNA fragments from a variety of sources, having a desirable gene sequence via appropriate vector.

Int J Genomics. 2016,  Role of Recombinant DNA Technology to Improve Life. Khan S. et al. doi: 10.1155/2016/2405954]]> 2020-10-26 06:07:53 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/860582599 ADVANTAGES juancarloflordeliza https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/860583062 Improved Medical Technology
Recombinant DNA technology has enhanced the efficiency of available detection systems in investigating and improving epidemiological information thereby increase disease awareness and make control and vaccination strategies easy.

Recombinant DNA technology has also contributed to health care in two important ways: production of pharmaceutically important proteins (biopharmaceuticals) and gene therapy for replacement of defective genes.

Enhanced Biological Characteristics
The outcomes of rDNA technology have a high degree of purity along with high specific activity. Some characteristics include greater yield, more resistance, improved nutrition and taste). Moreover since every preparation of the protein product is identical to the previous preparation batch-to-batch consistency is ensured.

Efficient Method
The genetic engineering utilizes modern tools and approaches such as molecular cloning and transformation, which are less time consuming and yield more reliable results.

Deb, R., Chakraborty, S., Sengar, G., & Bhanuprakash, V. (2016). Pros and Cons of Recombinant DNA Technology in Animal Diseases Diagnosis, Prevention and Control. Journal of Pure and Applied Microbiology; 10(1): 451-462.
https://microbiologyjournal.org/pros-and-cons-of-recombinant-dna-technology-in-animal-diseases-diagnosis-prevention-and-control/

Shinde, S.A.; Chavhan, S.A.; Sapkal, S.B.; Shrikhande, V.N. (2018). Recombinant DNA technology and its applications: A review. Int. J. Medipharm Res. 4, 79–88. [accessed Oct 29 2020]. ]]> 2020-10-26 06:08:08 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/860583062 DISADVANTAGES juancarloflordeliza https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/860583415 Effects on Biodiversity
The introduction of novel organisms into the environment could radically and detrimentally change ecosystems and could reduce biodiversity as a direct and indirect effect of modifying organisms.

The possibility of modified organisms released into the environment as bio-controlling agents becoming pathogenic to non-harmful organisms, pests developing resistance, and virus or bacteria becoming stronger and impervious to antibiotics.

Social Issues
Disease-producing organisms in rDNA experiments might develop infectious forms that could cause epidemics, and use of genetic engineering in biowarfare.

Ethical Issues
Creation of organisms containing altered genome and inheritance of modified genes rises the “playing God” argument against its use.

Patents on intellectual property (isolated genes, new gene constructs, recombinant cell types, GMOs) pose scientific problems. Limiting or preventing hold of patents for genes/tools reduce incentive for pursuing research. Genetic tests patenting create monopolies where one cannot get a second opinion if only one company holds the rights to conduct. Also, costs of a genetic test may be too high to afford.

Patra S, Andrew AA. Human, Social, and Environmental Impacts of Human Genetic Engineering. J Biomedical Sci. 2015, 4:2. doi:10.4172/2254-609X.100014. Retrieved October 27, 2020.

Rajakaruna SS, Taylor-Robinson AW. Application of recombinant DNA technology (genetically modified organisms) to the advancement of agriculture, medicine, bioremediation and biotechnology industries. J Appl Biotechnol Bioeng. 2016;1(3):78-80. DOI: 10.15406/jabb.2016.01.00013 Retrieved October 27, 2020

Science for Environment Policy (2016) Synthetic biology and Biodiversity, Future Brief 15. Bristol: Science Communication Unit, UWE; [cited 29 October 2020]. Available at: http://ec.europa.eu/science-environment-policy 

Snustad, D., & Simmons, M. (2011). Principles of Genetics (6th ed). Hoboken, NJ: John Wiley & Sons, Inc. Retrieved October 27, 2020

]]> 2020-10-26 06:08:19 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/860583415 RELEVANT RESEARCH ARTICLES juancarloflordeliza https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/860584409 Liu T, Li Z, Zhang Q, De Amorim Bernstein K, Lozano-Calderon S, Choy E, Hornicek FJ, Duan Z. Targeting ABCB1 (MDR1) in multi-drug resistant osteosarcoma cells using the CRISPR-Cas9 system to reverse drug resistance. Oncotarget. 2016 Dec 13;7(50):83502-83513. doi: 10.18632/oncotarget.13148. PMID: 27835872; PMCID: PMC5347784.

Porro, D., Sauer, M., Branduardi, P., & Mattanovich, D. (2005). Recombinant Protein Production in Yeasts. Methods in Molecular Biology, 824:329-58. doi: 10.1007/978-1-61779-433-9_17

Blake, A., Crockett, R., Essner, J., Hackett, P., and Nasevicius, A. (2018). Recombinant constructs and transgenic fluorescent ornamental fish therefrom. 20180064075 ]]> 2020-10-26 06:08:51 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/860584409 IN MEDICINE https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/863990286 Recombinant DNA technology has contributed to health care in two important ways: production of pharmaceutically important proteins (biopharmaceuticals) and gene therapy for replacement of defective genes.

Biopharmaceuticals
Recombinant DNA technology has been effectively used to produce various human proteins in microorganisms, such as insulin (the first commercial healthcare product derived) and growth hormone, used in the treatment of diseases. Also, the production of drugs against different viral, bacterial, and protozoan diseases are made through DNA technology.

Recombinant DNA technology has also proven important to the production of vaccines and protein therapies such as human insulin, interferon and human growth hormone. It is also used to produce clotting factors for treating hemophilia and in the development of gene therapy.

Gene therapy
Gene therapy aims to treat/cure/prevent disease by replacing a defective gene with a normal one using recombinant DNA technology.
In addition to these, personalized medicine, an emerging practice of medicine uses an individual's genetic profile to guide decisions made in regard to the prevention, diagnosis, and treatment of disease.

Diagnosis and Prediction of Diseases
The technology also forms the backbone of hepatitis and human immunodeficiency virus (HIV) diagnostic tests and for determining hereditary diseases.  This technology is also used  to predict the risk of developing a given disease and individual response to pharmacologic therapeutics—so called personalized medicine.

Rodwell, V. W., Bender, D. A., Botham, K. M., Kennelly, P. J., & Weil, P. A. (2018). Harper’s Illustrated Biochemistry (31st Ed.). USA: McGraw-Hill Education.

An Introduction to Ethical, Safety and Intellectual Property Rights Issues in Biotechnology( 2017)(Padma Nambisan) https://www.sciencedirect.com/topics/biochemistry-genetics-and-molecular-biology/recombinant-dna-technology
]]> 2020-10-27 00:37:38 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/863990286 IN AGRICULTURE https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/864265376 DNA recombinant technology is used to genetically modify crops with the purpose of improving both yield and resistance to plant pests or herbicides.

It is also through DNA recombinant technology that desired genes can be inserted into animals to increase the speed and range of breeding. It serves to help in the production of better farm animals.

Rajakaruna SS, Taylor-Robinson AW. Application of recombinant DNA technology (genetically modified organisms) to the advancement of agriculture, medicine, bioremediation and biotechnology industries. J Appl Biotechnol Bioeng. 2016;1(3):78-80. DOI: 10.15406/jabb.2016.01.00013
Shinde, S.A. et.al. (2018) Recombinant DNA Technology and its Applications: A Review. International Journal of MediPharm Research, 04(2), pp 79-88. Retrieved from: https://www.researchgate.net/publication/327111668_Recombinant_DNA_Technology_and_its_Applications_A_Review]]> 2020-10-27 03:17:23 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/864265376 WHAT IS A RECOMBINANT DNA (rDNA)? https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/864398932 It is a piece of DNA that has been created through laboratory methods of genetic recombination (e.g. molecular cloning) to combine genetic materials from multiple sources, creating sequences that would not otherwise be found in the genome.]]> 2020-10-27 04:46:36 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/864398932 GENETICALLY MODIFIED ORGANISMS (GMOs) https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/866199082 By utilizing the tools and techniques of genetic engineering it is possible to produce transgenic plants or the genetically modified plants. Many transgenic plants have been developed with better qualities like resistance to herbicides, insects or viruses or with expression of male sterility etc.

Genes for toxins that kill insects have been introduced in several species, including corn and cotton.

Anthony J.F. Griffiths, Encyclopedia Britanica Inc.  Recombinant DNA  (2018) retrieved October 27, 2020
https://www.britannica.com/science/recombinant-DNA-technology/In-vitro-mutagenesis

]]> 2020-10-27 15:33:31 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/866199082 RESTRICTION ENDONUCLEASES https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/867746529 Restriction endonucleases, also called restriction enzymes, are enzymes that bind to specific base, palindrome sequences in double-stranded DNA and catalyze the hydrolysis of phosphodiester bonds in both strands of the DNA, within or near the specific site. These enzymes provide the ability to generate a limited number of DNA fragments of predictable length and sequence from a DNA substrate.

Julin, Douglas (2014). Restriction Endonucleases. Molecular Life Sciences, 1-2. https://doi.org/10.1007/978-1-4614-6436-5_88-1 Retrieved October 27, 2020]]> 2020-10-27 22:16:23 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/867746529 https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872055059 2020-10-29 03:38:58 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872055059 GENE CLONING https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872063768 Gene cloning has proven particularly useful has been in mapping out the human genome, the creation of transgenic animals, and the development of insect-resistant crops.

Retrieved from: https://www.whatisbiotechnology.org/index.php/science/summary/rdna#:~:text=Recombinant%20DNA%20technology%20has%20also,the%20development%20of%20gene%20therapy.]]> 2020-10-29 03:46:18 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872063768 juancarloflordeliza https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872297843 2020-10-29 07:15:17 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872297843 PRACTICAL APPLICATIONS OF RECOMBINANT DNA TECHNOLOGY juancarloflordeliza https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872323134 2020-10-29 07:31:25 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872323134 juancarloflordeliza https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872328241 2020-10-29 07:34:51 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872328241 RECOMBINANT DNA TECHNOLOGY PROCESS: AN OVERVIEW juancarloflordeliza https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872332513 2020-10-29 07:37:25 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872332513 juancarloflordeliza https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872365492 2020-10-29 07:57:00 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872365492 juancarloflordeliza https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872369620 1. THE ENZYMES]]> 2020-10-29 07:59:28 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872369620 ADVANTAGES AND DISADVANTAGES OF DNA RECOMBINANT TECHNOLOGY juancarloflordeliza https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872392049 2020-10-29 08:14:44 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872392049 juancarloflordeliza https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872397328 2020-10-29 08:18:11 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872397328 juancarloflordeliza https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872399881 2020-10-29 08:19:49 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872399881 juancarloflordeliza https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872402613 2020-10-29 08:21:30 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872402613 IN FORENSIC SCIENCE https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872763859 The applications of rec DNA technology (or genetic engineering) in forensic sciences largely depend on the technique called DNA profiling or DNA fingerprinting. It enables us to identify any person by analyzing his hair roots Wood stains, serum, etc. DNA fingerprinting also helps to solve the problems of parentage and to identify the criminals.

Liu W., Yuan J. S., Stewart C. N., Jr  "GNN- Genetics and Genomics Timeline". www.genomenewsnetwork.org. Retrieved 29 October 2020 ]]> 2020-10-29 12:02:02 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872763859 TOOLS USED IN RECOMBINANT DNA TECHNOLOGY https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872776185 2020-10-29 12:07:49 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872776185 2. THE VECTORS https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872800128 Responsible  in carrying and integrating the desired gene. These form a very important part of the tools of recombinant DNA technology as they are the ultimate vehicles that carry forward the desired gene into the host organism. 
Plasmids and bacteriophages are the most common vectors in recombinant DNA technology.]]> 2020-10-29 12:17:53 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872800128 3. THE HOST ORGANISM https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872807711 into which the recombinant DNA is introduced. The host is the ultimate tool of recombinant DNA technology which takes in the vector engineered with the desired DNA with the help of the enzymes.

Shinde, S.A. et.al. (2018) Recombinant DNA Technology and its Applications: A Review. International Journal of MediPharm Research, 04(2), pages 83-84. Retrieved from: https://www.researchgate.net/publication/327111668_Recombinant_DNA_Technology_and_its_Applications_A_Review]]> 2020-10-29 12:21:03 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/872807711 anngelineelin https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/873454419 2020-10-29 14:58:37 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/873454419 IN THE ENVIRONMENT juancarloflordeliza https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/873560740 Genetic engineering is actively involved in developing biocatalysts for remediation of contaminated environments, and in the development of eco-friendly processes for biofuel production. Also, genetically modified organisms are used in cleaning up oil spills which is a major environmental hazard.

Retrieved from: https://www.alliedacademies.org/articles/benefits-of-genetic-engineering-for-human-welfare.pdf]]> 2020-10-29 15:20:54 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/873560740 anngelineelin https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/873632644 2020-10-29 15:35:25 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/873632644 anngelineelin https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/873666811 2020-10-29 15:42:02 UTC https://padlet.com/juancarloflordeliza/lug3lz1ymfbjys6p/wish/873666811