GBIO 70_05. Producing Genetically Modified Organisms PDF

11/01/2025 GBIO 60: Introduction to Biotechnology Producing...

11/01/2025 GBIO 60: Introduction to Biotechnology Producing Genetically TOPICS Discussion A. The Process of Genetic Engineering Modified Overview B. Genetic Modification of Microorganisms, Plants and Animals Organisms C. Cloning JAMAICA Q. ESPINELI SHAMDEE N. NAHAR 1 2 History of Biotechnology Biotechnology Innovations Innovations CATEGORIES OF MODERN BIOTECHNOLOGY PRINCIPLES OF BIOLOGY Genomics Cell Theory Proteomics Evolution (by natural selection) Metabolomics Gene Theory Homeostasis This course material is intended solely for the personal use of the student enrolled in GBIO 60. This course material is intended solely for the personal use of the student enrolled in GBIO 60. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. 3 4 1  11/01/2025 Techniques Used in Modern Biotechnology BY THE INTERCHANGE OF BY MUTATION GENETIC INFORMATION OR DNA BETWEEN LIKE which is a chemical deletion ORGANISMS or addition of one or more of the chemical parts of the DNA by sexual reproduction, and by molecule horizontal transfer in bacteria This course material is intended solely for the personal use of the student enrolled in GBIO 60. This course material is intended solely for the personal use of the student enrolled in GBIO 60. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. 5 6 ORGANISMAL MANIPULA T IO N Genetic manipulation of whole DEFINITION Genetic Engineering organisms CELLULAR MANIPULAT IO N Manipulation involve either cell fusion or the culture of cells and the regeneration of of Genetic whole plants from these cells; a semi-random or directed process Material and the changes can be more readily identified the formation of new combinations of heritable material by the insertion of nucleic acid molecules, MOLECULAR MANIPULA T IO N produced by whatever means outside the cell, manipulations of DNA and RNA; into any virus, bacterial plasmid or other vector system a directed control of the changes; so as to allow their incorporation into a host organism genetic engineering or in which they do not naturally occur, but in which they recombinant DNA technology are capable of continued propagation This course material is intended solely for the personal use of the student enrolled in GBIO 60. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. This course material is intended solely for the personal use of the student enrolled in GBIO 60. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. 7 8 2  11/01/2025 GOING IN-DEPTH CLONING process of making multiple, identical FO RMATIO N SP L IC IN G OF INTRODUC TION SE L E CTION OF DNA D N A IN TO OF vEC TORS OF N E WL Y copies of a particular INTO H OST FRAG ME N TS VE C TORS C ELLS ACQU IRE D piece of DNA DNA This course material is intended solely for the personal use of the student enrolled in GBIO 60. This course material is intended solely for the personal use of the student enrolled in GBIO 60. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. 9 10 1. CUTTING AND PASTING DNA Cut open the plasmid and "paste" in the 1. Cutting and gene. This process relies on restriction enzymes (which cut DNA) and DNA ligase pasting DNA Steps of (which joins DNA). RESTRICTION DNA 2. BACTERIAL TRANSFORMAT IO N AND SELECTION ENZYMES AND DNA LIGASE CLONING Insert the plasmid into bacteria. Use antibiotic selection to identify the A restriction enzyme is a DNA- bacteria that took up the plasmid. cutting enzyme that recognizes a specific target sequence and cuts 3. PROTEIN PRODUCTION DNA into two pieces at or near Grow up lots of plasmid-carrying bacteria that site. and use them as "factories" to make the DNA ligase seals gaps in the DNA protein. Harvest the protein from the backbone. bacteria and purify it. This course material is intended solely for the personal use of the student enrolled in GBIO 60. This course material is intended solely for the personal use of the student enrolled in GBIO 60. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. 11 12 3  11/01/2025 2. Bacterial 1. Cutting and transformation pasting DNA and selection IN DETAIL Building a recombinant plasmid PLASMIDS AND 1. Separately digest (cut) the gene ANTIBIOTIC fragment and the plasmid with EcoRI. RESISTANCE GENE 2. Take the gene fragment and the Plasmids and other DNA can be linearized (opened-up) plasmid introduced into bacteria in a and combine them along with DNA process called transformation. ligase. A plasmid typically contains an 3. Once they are joined by ligase, antibiotic resistance gene, which the fragments become a single allows bacteria to survive in the piece of unbroken DNA. presence of a specific antibiotic. This course material is intended solely for the personal use of the student enrolled in GBIO 60. This course material is intended solely for the personal use of the student enrolled in GBIO 60. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. 13 14 2. Bacterial 3. Protein transformation and production selection P O S S I B I LI TY 1 : IN DETAIL BACTE RIA = PLASM ID FACTORIE S P O S S I B I LI TY 2 : BACTE RIA = PROTE IN FACTORIE S The bacteria serve as miniature “factories," churning out large amounts of protein. This course material is intended solely for the personal use of the student enrolled in GBIO 60. This course material is intended solely for the personal use of the student enrolled in GBIO 60. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. 15 16 4  11/01/2025 REPRODUCTIVE CLONING METHODS primarily used to produce genetically modified animals capable of passing along the new PRODUCTION OF GENETICALLY characteristics to its offspring SOMA TIC CELL CLONING MODIFIED involves the cloning of body cells, or somatic cells, to regenerate a whole ORGANISMS organism STEM CELL CLONING Stem cells have the natural ability to form tissues, organs, and whole organisms. This course material is intended solely for the personal use of the student enrolled in GBIO 60. This course material is intended solely for the personal use of the student enrolled in GBIO 60. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. 17 18 Transformation Transduction GENE INSERTION METHOD GENE INSERTION METHOD viruses that infect bacteria move a bacterium takes in DNA from its short pieces of chromosomal DNA environment, often DNA that's been from one bacterium to another "by shed by other bacteria. accident." This course material is intended solely for the personal use of the student enrolled in GBIO 60. This course material is intended solely for the personal use of the student enrolled in GBIO 60. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. 19 20 5  11/01/2025 Conjugation Transfection GENE INSERTION METHOD DNA is transferred from one GENE INSERTION METHOD bacterium to another. After the donor cell pulls itself close Heat shock to the recipient using a structure Electroporation called a pilus, DNA is transferred between cells. Chemoporation In most cases, this DNA is in the form of a plasmid. This course material is intended solely for the personal use of the student enrolled in GBIO 60. This course material is intended solely for the personal use of the student enrolled in GBIO 60. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. 21 22 Other Means Transgenic Plant GENE INSERTION METHOD AGROBACTERIUM-MEDIATED Bioballistics Ti Plasmid Microinjection Transfection Hybridoma formation Liposome fusion This course material is intended solely for the personal use of the student enrolled in GBIO 60. This course material is intended solely for the personal use of the student enrolled in GBIO 60. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. 23 24 6  11/01/2025 Transgenic Stem Cell Cloning Animal Therapeutic REPRODUCTIVECLONING Nuclear Programming Dolly This course material is intended solely for the personal use of the student enrolled in GBIO 60. This course material is intended solely for the personal use of the student enrolled in GBIO 60. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. 25 26 Alberts, B., Johnson, A., Lewis, J., Raff, M., Roberts, K., & Walter, P. (2002). Molecular biology of the cell. New York: Garland Science. Clark, D. P., & Pazdernik, N. J. (2009). Applying the Genetic Revolution. Uses of DNA cloning Herren, R.V. (2013). Introduction to Biotechnology: An Agricultural Revolution (2nd ed.). Asia: Delmar, Cengage Learning Shmaefsky, B. (2006). Biotechnology 101. Greenwood Publishing Group. Smith, J. E. (2009). A concise history of biotechnology–Some key determinants. Biotechnology, 1, 321-360. https://www.khanacademy.org/science/high-school-biology/hs-molecular-genetics/hs-biotechnology/a/hs- biotechnology-review https://www.khanacademy.org/science/biology/biotech-dna-technology/dna-cloning-tutorial/a/restriction- enzymes-dna-ligase https://www.khanacademy.org/science/biology/biotech-dna-technology/dna-cloning-tutorial/a/bacterial- transformation-selection https://www.khanacademy.org/science/ap-biology/gene-expression-and-regulation/mutations- Biopharmaceuticals Gene therapy Gene analysis References ap/a/genetic-variation-in-prokaryotes https://www.canva.com This course material is intended solely for the personal use of the student enrolled in GBIO 60. No part of this course material can be reproduced, reposted, or replicated in any form without permission from the instructors. 27 28 7