...The Transformation of Escherichia coli With pGalTM OBJECTIVE: To develop an understanding of bacterial transformation by plasmid DNA, and determine the transformation efficiency of a bacterial sample. INTRODUCTION: Transformation is an important biological mechanism for generating genetic diversity in the natural world. It is also one of the most used tools used in modern biotechnology. It has allowed for a lot of genetic engineering advances. Scientists can design and build DNA containing genes they want and then transfer these genes into bacterial cells that they can get to express those genes. This is a very basic technique that is used on a daily basis in a molecular biological laboratory. This is based on the natural function of a plasmid: to transfer genetic information vital to the survival of the bacteria. Transformation is an example of horizontal gene transfer. There are three mechanisms for horizontal gene transfer in bacteria. Conjugation is the transfer of DNA from one cell to another through conjugation pili. Transduction is the transfer of genes between bacterial cells using a bacteriophage (a type of virus). Transformation is the ability of cells to take up freely floating DNA found in the environment. Bacterial cells that are able to take up free-floating DNA from the environment are called competent cells. Bacteria are not always competent. When growth conditions are optimal, most bacteria cannot uptake DNA. In most cases, in response to some stimuli...
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...Understanding The Transformation Of E. Coli Bacteria Using Plasmid DNA pGLO To Trigger Resistance To Ampicillin. Author: Nick Delagarza Section: Tuesday 11:30, Room 121 TA: Huy D. Vu ABSTRACT – Transformation is the uptake of naked DNA in the environment by a recipient bacterium. The purpose of this experiment is to perform genetic transformations of E. Coli bacteria using plasmid DNA in forming resistance to Ampicillin. Escherichia coli is a bacterium is found in the human colon and in many areas of the environment. The Foreign DNA used in this experiment contains the gene AmpR, which makes the transformed bacteria Ampicillin resistant. The plasmid also contains a gene for GFP, which would allow the bacteria to glow under UV light. The hypothesis in this lab states that if the E. Coli cell is accompanied by the pGLO plasmid, the cell will grow in environment containing Ampicillin, but if only containing Ampicillin without the plasmid growth would be inhibited. The other part of the hypothesis explains that in an environment containing Ampicillin, Arabinose, and the plasmid, the bacteria would both grow and glow. At the end of the experiment, the results proved that E. Coli could be transformed using foreign plasmid DNA to form Ampicillin resistance. INTORDUCTION Genetic Engineering is important in several fields of biotechnology and is bigger than ever in improving human insulin. There are many examples today on the use of genetic engineering and biotechnology. First and...
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...As we have sped into the 21st century, we have run into the issue of keeping up food supply with the demands of the growing population. To keep up with these new demands, large food companies have enlisted the help of scientists to engineer our foods to make them easier, faster, and cheaper to grow. Unfortunately, using Genetically Modified Organisms (GMOs) is bad not only for the health of us humans, but also the health of our environments. I believe that we should stop using genetically modified foods so that we can avoid liver disease, kidney disease and an increased risk of cancer in humans, and stop the rise of antibiotic resistance in animals and on crops. Throughout the past several decades, as we have funded the genetic engineering of our food, the higher ups of the food companies have turned a blind eye to the health risks that consuming engineered food have to us, and have conveniently forgotten to inform the everyday consumer of these risks. In “Evidence Grows of Harmful Effects of GMOs on Human Health” by Helke Ferrie, who has a MA in Physical Anthropology and has spent much time working with the World Health Organization, states that according to Canadian research “93% of all fetuses and their mothers tested were found to be contaminated with toxins associated with GM foods” (Ferrie 2) and that there is proof “GM soy products cause pre-cancerous changes to the ovaries and uterus of animals that consume them” (Ferrie 2). To begin with, the most important stage of...
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...Bacteria |I | |INTRODUCTION | Bacteria, one-celled organisms visible only through a microscope. Bacteria live all around us and within us. The air is filled with bacteria, and they have even entered outer space in spacecraft. Bacteria live in the deepest parts of the ocean and deep within Earth. They are in the soil, in our food, and on plants and animals. Even our bodies are home to many different kinds of bacteria. Our lives are closely intertwined with theirs, and the health of our planet depends very much on their activities. Bacterial cells are so small that scientists measure them in units called micrometers (µm). One micrometer equals a millionth of a meter (0.0000001 m or about 0.000039 in), and an average bacterium is about one micrometer long. Hundreds of thousands of bacteria would fit on a rounded dot made by a pencil. Bacteria lack a true nucleus, a feature that distinguishes them from plant and animal cells. In plants and animals the saclike nucleus carries genetic material in the form of deoxyribonucleic acid (DNA). Bacteria also have DNA but it floats within the cell, usually in a loop or coil. A tough but resilient protective shell surrounds the bacterial cell. Biologists classify all life forms as either prokaryotes or eukaryotes. Prokaryotes are simple, single-celled organisms like bacteria. They lack a defined nucleus of the sort found in plant and animal cells. More complex organisms, including all plants and animals, whose cells have a...
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... Name partner: Marieke RoosStudent Numbers: 1215671/4228073 Practicum assistants: Brijith Thomas,Room: 1 Joanna Pawlak, Lara van der Woude, Date: 12/12/2014 Valerie SelsEmail: saburo_aikini441@yahoo.com | Lab Report DNA: Plasmids and Nucleases 1. Abstract The goal of this practicum was to isolate plasmid DNA from Escherichia coli (E. coli), to identify it, to prove that the plasmid is circular and double-stranded and to give bacterial cells new genetic properties via transformation. An unknown plasmid S was isolated from the bacterial stain Escherichia coli (E. coli). Then its purity was determined by calculating the ratio A260/A280. After that, the unknown plasmid S was identified, followed by the determination of its antibiotic resistance gene. Subsequently, a new culture of E. coli DH5α was transformed by using the isolated plasmids where only the transformed cells survived. Finally, the plasmid-DNA was treated with exonuclease V to prove the circular and double-stranded property of plasmid. The isolated plasmid S was identified to be pCTB2 which was found to be have two forms: (I) relaxed & (II) supercoiled. The ratio A260/A280 gave the value of 1.65 indicating the presence of proteins (RNA/contaminated DNA). Furthermore, the plasmids were partially digested with restriction enzymes...
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...The evolution of multi drug resistant strain shigella sonnei from its ancestor E. Coli and predecessor S. Flexneri and its spread to Vietnam Introduction Shigella is a genus of Gram-negative, facultative anaerobic, nonspore-forming, nonmotile, rod-shaped bacteria. The genus is named after Kiyoshi Shiga, who first discovered it in 1897. The causative agent of human shigellosis, Shigella causes disease in primates, and is the only naturally found in humans and apes. Shigella is one of the leading bacterial causes of diarrhea worldwide and it is suggested that Shigella causes about 90 million cases of severe dysentery, with at least 100,000 of these resulting in death each year. Shigella originates from a human-adapted E. coli that has gained the ability to invade the human gut mucosa, spreading through fecal-oral transmission via contaminated food or water. Depending on age and condition of the host, fewer than 100 bacterial cells can be enough to cause an infection. The infection results in the destruction of the epithelial cells of the intestinal mucosa. Some strains produce the enterotoxin shiga toxin, which is similar to the verotoxin of E. coli. The species invades the host through the cells in the gut epithelia of the small intestine. Shigella uses a type-III secretion system, which acts as a biological syringe to translocate toxic effector proteins to the target human cell. The effector proteins can alter the metabolism of the target cell, making it easier for intracellular...
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...NINTH EDITION Burton’s MICROBIOLOGY FOR THE HEALTH SCIENCES Paul G. Engelkirk, PhD, MT(ASCP), SM(AAM) Biomedical Educational Services (Biomed Ed) Belton, Texas Adjunct Faculty, Biology Department Temple College, Temple, TX Janet Duben-Engelkirk, EdD, MT(ASCP) Biomedical Educational Services (Biomed Ed) Belton, Texas Adjunct Faculty, Biotechnology Department Temple College, Temple, TX Acquisitions Editor: David B. Troy Product Manager: John Larkin Managing Editor: Laura S. Horowitz, Hearthside Publishing Services Marketing Manager: Allison Powell Designer: Steve Druding Compositor: Maryland Composition/Absolute Service Inc. Ninth Edition Copyright © 2011 Lippincott Williams & Wilkins, a Wolters Kluwer business © 2007 Lippincott Williams & Wilkins, © 2004 Lippincott Williams & Wilkins, © 2000 Lippincott Williams & Wilkins, © 1996 Lippincott-Raven, © 1992, 1988, 1983, 1979 JB Lippincott Co. 351 West Camden Street Baltimore, MD 21201 Printed in the People’s Republic of China All rights reserved. This book is protected by copyright. No part of this book may be reproduced or transmitted in any form or by any means, including as photocopies or scanned-in or other electronic copies, or utilized by any information storage and retrieval system without written permission from the copyright owner, except for brief quotations embodied in critical articles and reviews. Materials appearing in this book prepared by individuals as part of their official duties as U.S. government employees...
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...Membrane lipids feature ether linkages (1) More stable under extreme environmental conditions c) Archaea do not have peptidoglycan cell walls (1) Have outer coverings made of protein instead B. Bacteria 1. Prokaryotic cell structure 2. Membrane lipids are formed with ester linkages 3. Cell walls can contain peptidoglycan C. Domain Archaea Was Ancestral to Domain Eukarya Fig. 27.2 Bacteria and Archaea !2 1. DNA with histone proteins (shared with Eukarya) (1) Archaea and Eukarya share over 30 ribosomal proteins that are not present in bacteria (2) Archaeal and Eukaryotic RNA polymerases are closely related 2. Ether-linked cell membranes a) Ester bonds are in bacteria and eukaryotes b) Ether bonds are unique to archaea (1) Ether bonds are resistant to damage by heat and other extreme conditions (a) Explains why extremophiles are able to exist c) Archaea also use isoprene chains instead of fatty acid chains in making membranes 3. Extremophiles a) Organisms that occur primarily in extreme habitats 4. Hypothermophiles a) An organism that thrives in extremely hot environments b) Methanopyrus (1) Grows best at deep sea thermal vent sites (a) Where the temperature is 98 degrees celsius Both are Methanopyrus (b) It actually can not grow when the temperature is less than 84 degrees celsius (2) In the phylum Euryarchaeota 5. High temperatures and low pH a) Sulfolobus (1) From sulfur hot springs with a pH of 3 or lower (2) In the phylum Crenarchaeota 6. Halophiles a) Species able to grow...
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...INTRODUCTION Infection remains the main cause of morbidity and mortality in man, particularly in developing areas where it is associated with poverty and overcrowding.In the developed world increasing prosperity, universal immunization and antibiotics have reduced the prevalence of infectious disease. However, antibiotic-resistant strains of microorganisms and diseases such as human immunodeficiency virus (HIV) infection, variant Creutzfeldt–Jakob disease ,avian influenza and severe acute respiratory syndrome(SARS) have emerged. In the developing world successes such as the eradication of smallpox have been balanced or outweighed by the new plagues. Infectious diseases cause nearly 25% of all human deaths. Two billion people one-third of the world’s population are infected with tuberculosis (TB),500 million people catch malaria every year, and 200 million are infected with schistosomiasis. 39.5 million people are living with HIV/AIDS, with 4.3 million new HIV infections in 2006 (65% in sub-Saharan Africa). The eight Millennium Development Goals (MDG) hope to be achieved by 2015. These are: eradicating extreme poverty and hunger, universal primary education, gender equality and empowerment of women, reduction in child mortality,improvement in maternal health, combating HIV/AIDS,malaria and other diseases, ensuring environment sustainability and developing global partnership for development. DEFINITION INFECTION An infection is the entry and multiplication of an infections...
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...MSc Biotechnology, Bioprocessing & Business Management 2014/15 Module name and number Fundamentals of Biotechnology: BS934 Assignment title Antimicrobials Student Number 1464986 Word Count 2470 The article will initially cover conventional antibiotics and how their abuse led to antibiotic resistance in bacteria. Alternative antimicrobials will be reviewed by taking into consideration their mechanisms of action and how they can represent a viable option to address the issue of antibiotic resistance. Antimicrobial Background ------------------------------------------------- Figure 1: Structure of penicillin and β-lactam ring (Adapted from LNLS, 2013) ------------------------------------------------- Figure 1: Structure of penicillin and β-lactam ring (Adapted from LNLS, 2013) Antimicrobials are substances that kill or suppress microorganisms such as bacteria, fungi and viruses (EPA, 2014). The first evidence of antimicrobials dates back to the Egyptians who used mouldy bread to treat surface infections (Wainwright, 1989). At the beginning of the 20th century, Paul Ehrlich synthesized the first antimicrobial agent known as Salvarsan, a drug used for treating syphilis (Aminov, 2010). Successively Alexander Fleming discovered penicillin by noticing that the growth of Staphylococcus aureus was inhibited in presence of the mold produced by Penicillium fungi. This led to the conclusion that microorganisms produce substances to limit...
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...will be faced daily with the concepts of microbial disease and antimicrobial therapy. Microbiology is one of the few courses where much of the "minutia" is regularly used by the practicing physician. This book attempts to facilitate the learning of microbiology by presenting the information in a clear and entertaining manner brimming with memory aids. Our approach has been to: 4) Create a conceptual, organized approach to the organisms studied so the student relies less on memory and more on logical pathophysiology. The text has been updated to include current information on rapidly developing topics, such as HIV and AIDS (vaccine efforts and all the new anti-HIV medications), Ebola virus, Hantavirus, E. coli outbreaks, Mad Cow Disease, and brand-new antimicrobial antibiotics. The mnemonics and cartoons in this book do not intend disrespect for any particular patient population or racial or ethnic group but are solely presented as memory devices to assist in the learning of a complex and important medical subject. We welcome suggestions for future editions. 1) Write in a conversational style for rapid assimilation. 2) Include numerous figures serving as "visual memory tools" and summary charts at the end of each chapter. These can be used for "cram sessions" after the concepts have been studied in the text. 3) Concentrate more on clinical and infectious disease issues that are both interesting and vital to the actual practice of medicine. MARK GLADWIN, MD BILL TRATTLER, MD...
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...1. The sudden unexpected occurrence of a disease in a given population: a. Outbreak b. Zoonotic c. Endemic d. Sporadic 2. A causative agent of peptic ulcer a. Escherichia coli b. Bordetella pertusis c. Shigella dysenteriae d. Helicobacter pylori 3. The “four o’clock habit” promoted by the Department of health is designed to prevent the spread of: a. Dengue fever b. Tuberculosis c. Malnutrition d.malaria 4. The first drug available for HIV: a. ZDU b. RMP c. TMP d. CMC 5. An immediate hypersensitivity reaction following exposure of a sensitized individual to the appropriate antigen: a. Hay fever b. Anaphylaxis c. Asthma d. Desensitation 6. A thin proteinaceous appendage necessary for bacterial conjugation: a. Cilium b. Trichome c. Flagellum d. Pilus 7. The use of nitrite as preservative for food is discouraged because: a. It can discolour the meat b. It can react with amines to form carcinogenic nitrosamines c. It decomposed to nitric acid which can react with heme pigments d. It is not readily available 8. The disinfectant of choice for municipal water supply: a. Lysol b. Chlorine c. Ozone d. Reverse osmosis 9. An object that is able to harbour and transmit microorganisms: a. Mite b. Fomite c. Arthropod d. Vector 10. Administration of a toxoid cenfers: a. Naturally acquired active immunity b. Naturally acquired passive immunity c. Artificially acquired active immunity d. Artificially...
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...while on the contrary remaining “dormant” when not in need. This has been accomplished at the molecular level by configuring clusters of genes together on the genome into operons that elicit a processive response in the presence of a specific metabolite. The Lac operon is responsible for the cleaving of the disaccharide lactose into two products. A myriad of components control the expression of the Lac operon when two conditions are met. First, the substrate, lactose, must be present. Second, no better substrate for example, glucose, is present (2). The three structural genes in the Lac operon are lacZ, lacY, and lacA. The gene lacZ encodes the tetramer, ß-galactosidase, which is responsible for hydrolyzing the ß-1,4 glycosidic linkage between galactose and glucose in lactose. The transport of lactose into the cell via the enzyme lactose permease is encoded by the gene lacY. The lacA gene encodes the enzyme, galactoside transacetylase, a trimer that transfers an acetyl group from acetyl-CoA to galactosides. Activation of these genes is dependent on the activity of a promoter and three operators based on the nutritional and environmental conditions available to the cell. The lac operon is a negatively controlled inducible operon that utilizes the product of the regulator gene lacI, to repress RNA polymerase from transcribing the lacZYA genes. The three operators involved in the lac operon: O1, O2, and O3 serve as binding sites for lacI and when bound repression is exhibited. Originally...
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...Aacknowledgement First of all I like to thank God almighty who authorise me to research on this topic. I submit my sincere thanks to my supervisor Medhat Khattar for his valuable presence, time, effort, guidance and help to complete this dissertation. My dissertation would not have been completed without the help of lab technicians Nick and Suzy, I am extremely grateful for their help, suggestions and encouragement. I might want to thank my family for impacting in me a comprehension for the significance of education and an appreciation for diligent work. I extraordinarily value the majority of the penances that were made so as to realize the open doors that I have gotten, and it is my trusts that this proposition embodies what I have realized. Much obliged to you for your dedication, bolster, and affection. I might likewise want to thank my grandparents for the numerous hours of math mentoring as a youngster. In spite of the fact that it may have appeared to be inconsequential, it was the premise for my prosperity and the establishment of my hobbies in Designing. I might want to devote this proposition to my family, without whom I would not be seeking after a profession with an instruction from my university undergraduate days. I also thank my supervisor’s effort and good work channelled towards making me a better microbiologist in the world. I sincerely extend my thanks all concerned people who together with me in this regard. Table of Contents I Declaration......
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...positive attributes. However, GMOs also have adverse effects on animals, the environment and the health of the consumer. In order to understand why GMOs have these effects, it is important to know exactly what a GMO is and how these differ from the normal breeding farmers have done for years. The Free Dictionary defines a GMO as “An organism whose genetic characteristics have been altered using the techniques of genetic engineering.” It goes on to explain that modern science has the ability to modify the genetic makeup of plants and animals. Scientists can even transfer genes from one species to another. http://www.thefreedictionary.com/genetically+modified+organism Advocates of genetically modified organisms say that the use of the gmos could end world hunger they also say we could create plants and animals that are resistant to disease and tolerant to pesticides. They can also engineer plants to be drought resistant. They argue these changes would stop famine because the plants could naturally resist. The proponents use pathos to convince people they have the answer to world hunger. They show you pictures starving people around the world and tell you their “improved” seeds make hunger go away. Perhaps it could but the studies do not prove that yet. In fact, the only feeding study done with humans showed that GMOs survived inside the stomach of the people eating GMO food. This does not mean it is safe for our consumption. No other follow-up studies were done to determine the...
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