GMO, genetic engineering,geniticaly modified organisms

genetically modified organisms

1. Ways in which these plasmids may be introduced into host organisms: ❖ Biolistics . In this technique, a “gene gun” is used to fire DNA-coated pellets on plant tissues. Cells that survive the bombardment, and are able to take up the expression plasmid coated pellets and acquire the ability to express the designed protein.

2. Ways in which these plasmids may be introduced into host organisms: ❖ Plasmid insertion by Heat Shock Treatment. Heat Shock Treatment is a process used to transfer plasmid DNA into bacteria. The target cells are pre-treated before the procedure to increase the pore sizes of their plasma membranes. This pretreatment (usually with CaCl2) is said to make the cells “competent” for accepting the plasmid DNA. After the cells are made competent, they are incubated with the desired plasmid at about 4°C for about 30min.

3. Ways in which these plasmids may be introduced into host organisms: The plasmids concentrate near the cells during this time. Afterwards, a “Heat Shock” is done on the plasmid-cell solution by incubating it at 42°C for 1 minute then back to 4°C for 2 minutes. The rapid rise and drop of temperature is believed to increase and decrease the pore sizes in the membrane. The plasmid DNA near the membrane surface are taken into the cells by this process. The cells that took up the plasmids acquire new traits and are said to be “transformed”.

4. Electroporation. This technique follows a similar methodology as Heat Shock Treatment, but, the expansion of the membrane pores is done through an electric “shock”. This method is commonly used for insertion of genes into mammalian cells .

5. Some methods are: • Selection of plasmid DNA containing cells • Selection of transformed cells with the desired gene • PCR detection of plasmid DNA • Genetically Modified Organisms (GMOs)

6. Genetically modified organisms (GMOs) - are organisms whose genetic material has been synthetically manipulated in a laboratory through genetic engineering. GMOs refer broadly to organisms that are produced when selected individual genes are transferred from a given donor organism into another target organism, typically conferring desired properties to the new organism. 

7. GMOs can include plants, animals, and enzymes. Some GMOs have been approved by regulatory agencies for commercial production and consumption, while others are currently undergoing regulatory evaluation. Practically, all commercial GMOs are engineered to withstand direct applications of herbicide and/ or to produce an insecticide.

8. O ther GMOs are in experimental stages and confined to scientific laboratory research. According to the United States Department of Agriculture (USDA), by 2012, 93% of soybeans, 94% of cotton, and 88% of corn grown in the U.S. were genetically modified (Center for Eco genetics and Environmental Health).

9. Classical Breeding o It focuses on the mating of organisms with desirable characteristics. o May develop new plant varieties in the selection process and seek to achieve expression of genetic material present within a species.

10. o It employs processes that occur in nature, i.e., sexual and asexual reproduction. o The product emphasizes certain characteristics and not new for the species, which have been present for millennia within the species' genetic potential (Hansen,2000)

11. Gene Cloning A process by which large quantities of a specific, desired gene or section of DNA may be cloned or copied once the desired DNA has been isolated. 1. The gene or DNA that is desired is isolated using restriction enzymes. 2. Both the desired gene and a plasmid are treated with the same restriction enzyme to produce identical sticky ends. 3. The DNAs from both sources are mixed together and treated with the enzyme DNA ligase to splice them together.

12. Gene Cloning 4. Recombinant DNA, with the plasmid containing the added DNA or gene, has been formed. 5. The recombinant plasmids are added to a culture of bacterial cells. Under the right conditions, some of the bacteria will take in the plasmid from the solution during a process known as transformation. 6. As the bacterial cells reproduce (by mitosis), the recombinant plasmid is copied. Soon, there will be millions of bacteria containing the recombinant plasmid with its gene. 7. The introduced gene can begin producing its protein via transcription and introduced translation.

13. Applications of Recombinant DNA

14. E xamples of modified traits using cloned genes and their applications

15. Recombinant DNA Recombinant DNA technology is a technique that changes the phenotype of an organism (host) when a genetically altered vector is introduced and integrated into the genome of the organism. So, the process involves the introduction of a foreign piece of DNA structure into the genome, which contains our gene of interest. 

16. Recombinant DNA This gene that is introduced is the recombinant gene, and the technique is called recombinant DNA technology. Inserting the desired gene into the genome of the host is not as easy as it sounds.

17. Recombinant DNA It involves the selection of the desired gene for administration into the host, followed by a selection of the perfect vector with which the gene has to be integrated and recombinant DNA formed. This recombinant DNA then has to be introduced into the host. And at last, it has to be maintained in the host and carried forward to the offspring (Shinde et.al. 2018).

18. Recombinant DNA The primary tools of recombinant DNA technology are bacterial enzymes called restriction enzymes . Each enzyme recognizes a short, specific nucleotide sequence in DNA molecules and cuts the molecules' backbones at that sequence.

19. Recombinant DNA The result is a set of double-stranded DNA fragments with single-stranded ends, called " sticky ends ." Sticky ends are not really sticky; however, the bases on the sticky ends form base pairs with the complementary bases on other DNA molecules. Thus, the sticky ends of DNA fragments can be used to join DNA pieces originating from different sources.

20. Recombinant DNA

21. Applications of Recombinant DNA Technology (Adapted from Shinde, et al., 2018 1. Production of Transgenic Plants By utilizing the tools and techniques of genetic engineering, it is possible to produce transgenic plants or genetically modified plants. Many transgenic plants have been developed with better qualities like resistance to herbicides, insects, or viruses or with the expression of male sterility, etc.

22. Applications of Recombinant DNA Technology (Adapted from Shinde, et al., 2018 2. Production of Transgenic Animals By the use of rec DNA technology, desired genes can be inserted into the animal so as to produce the transgenic animal. The method of rec DNA technology aids the animal breeders to increase the speed and range of selective breeding in the case of animals. It helps for the production of better farm animals to ensure more commercial benefits. Another commercially important use of transgenic animals is the production of specific proteins and pharmaceutical compounds. Transgenic animals also contribute to studying the gene functions in different animal species. Biotechnologists have successfully produced transgenic pigs, sheep, rats, and cattle.

23. Applications of Recombinant DNA Technology (Adapted from Shinde, et al., 2018 3. Production of Hormones By the advent of techniques of rec DNA technology, bacterial cells like E.coli are utilized for the production of different fine chemicals like insulin, somatostatin, somatotropin, and endorphin. Human Insulin Hormone, i.e., Humulin, is the first therapeutic product that was produced by the application of rec DNA technology. .

24. Applications of Recombinant DNA Technology (Adapted from Shinde, et al., 2018 4. Production of Vaccines Vaccines are the chemical preparations containing a pathogen in an attenuated (or weakened) or inactive state that may be given to human beings or animals to confer immunity to infection. A number of vaccines have been synthesized biologically through recDNA technology; these vaccines are effective against numerous serious diseases caused by bacteria, viruses, or protozoa. These include vaccines for polio, malaria, cholera, hepatitis, rabies, smallpox, etc. The generation of DNA vaccines has revolutionized the approach to the treatment of infectious diseases. DNA-vaccine is the preparation that contains a gene encoding an immunogenic protein from the concerned pathogen . .

25. Applications of Recombinant DNA Technology (Adapted from Shinde, et al., 2018 5. Biosynthesis of Interferon Interferons are the glycoproteins that are produced in very minute amounts by the virus-infected cells. Interferons have antiviral and even anti-cancerous properties. By the recDNA technology method, the gene of human fibroblasts (which produce interferon's in human beings) is inserted into the bacterial plasmid. These genetically engineered bacteria are cloned and cultured so that the gene is expressed and the interferons are produced in relatively high quantities. This interferon, so produced, is then extracted and purified.

26. Applications of Recombinant DNA Technology (Adapted from Shinde, et al., 2018 6. Production of Antibiotics Antibiotics produced by microorganisms are very effective against different viral, bacterial, or protozoan diseases. Some important antibiotics are tetracycline, penicillin, streptomycin, novobiocin, bacitracin, etc. The recDNA technology helps in increasing the production of antibiotics by improving the microbial strains through modification of genetic characteristics.

27. Applications of Recombinant DNA Technology (Adapted from Shinde, et al., 2018 7. Production of Commercially Important Chemicals Various commercially important chemicals can be produced more efficiently by utilizing the methods of rec DNA technology. A few of them are the alcohols and alcoholic beverages obtained through fermentation, organic acids like citric acid, acetic acid, etc., and vitamins produced by microorganisms

28. Applications of Recombinant DNA Technology (Adapted from Shinde, et al., 2018 8. Application in Enzyme Engineering As we know that the enzymes are encoded by genes, so if there are changes in a gene, then definitely the enzyme structure also changes. Enzyme engineering utilizes the same fact and can be explained as the modification of an enzyme structure by inducing alterations in the genes which encode for that particular enzyme.

29. Applications of Recombinant DNA Technology (Adapted from Shinde, et al., 2018 9. Prevention and Diagnosis of Diseases Genetic engineering methods and techniques have greatly solved the problem of conventional methods for the diagnosis of diseases. It also provides methods for the prevention of a number of diseases like AIDS, cholera, etc. Monoclonal antibodies are useful tools for disease diagnosis. Monoclonal antibodies are produced by using the technique called hybridoma technology.

30. Applications of Recombinant DNA Technology (Adapted from Shinde, et al., 2018 10. Gene Therapy Gene therapy is undoubtedly the most beneficial area of genetic engineering for human beings. It involves the delivery of specific genes into the human body to correct the diseases. Thus, it is the treatment of diseases by transfer and expression of a gene into the patients' cells so as to ensure the restoration of a normal cellular activity