Exam 2-Scott – Flashcards
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Selective Media |
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Inhibits growth of one bacteria over another. EG: McConkey is selective for gram negative and Mannitol Salt is selective for gram positive |
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Enrichment Media |
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All bacteria types can grow but only one will outgrow all the rest because of nutrients. EG: blood agar, fastidious microorganisms |
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Differential Media |
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Distinguish one microorganism type from another growing on the same media. McConkey is differential for lactose fermentation and mannitol salt agar is differential for mannitol fermentation |
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Obligate anaerobe |
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Microorganisms that live and grow in the absence of molecular oxygen. They gather at the bottom of the test tube to avoid oxygen. They do not have enzymes, and lack an electron transport chain. That is why we can't use oxygen to ...? These form endospores and are also thermophiles |
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Facultative anaerobe |
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These are not obligate. The anaerobe does not need oxygen to grow. Lack of oxygen does not hurt them. they can be found all along the test tube. They are adapting to variations. Have both enzymes (superoxide dismutase and catalase). Eg; E. coli and S. aureus |
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Microaerophile |
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Require oxygen but at a low concentration. So they will gather towards upper end of test tube, but not at bottom. Have little enzymes. Eg: Heliobacter Pylori - also a neutrophile and makes an enzyme called Urease which changes the pH from 3 to 7 so it can survive in the stomach. Causes ulcer. |
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Obligate aerobe |
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Oxygen-needing bacteria. Gather at the top of test tube in order to absorb max amount of O2. Has enzymes. Survival depends on their ability to detoxify O2 and successful gene expression. Eg: Bordetella Pertussis (whooping cough). |
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Aerotolerant Bacteria |
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Are not affected at all by oxygen. The are evenly spread along the test tube. They only contain the enzyme superoxide dismutase (SOD) Use Peroxidase to break down peroxide and superoxide. |
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Describe the natural antioxidants in our bodies |
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1. Superoxide dismutase (SOD) - removes the free radical from oxygen making o2 and H2O2 (hydrogen peroxide). (or the reverse?) 2. Catalase - Hurns H2O2 into O2 and H2O so now it can safely go into the body. |
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Hemolysins |
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Enzymes that break down red blood cells. Red blood cells carry oxygen. |
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Lag phase of growth curve |
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Early phase of bacteria population grown during which no signs of growth occur as bacteria are adjusting to their environment. |
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Log phase of growth curve |
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maximum rate of cell division during which growth is geometric in rate of increase. Also called exponential growth |
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Death phase of growth curve |
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End of the cell growth due to lack of nutrition, depletion of environment, accumulation of wastes. Population of cells begins to die. More cells are dying than dividing. |
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Stationary phase |
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survival mode, balance is equal between cells dividing and cells dying |
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CFU - Colony-forming unit |
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A measure of viable bacterial numbers. Unlike direct microscopic counts where all cells, dead and living, are counted, CFU measures viable cells. Results are given as CFU/ml for liquids, CFU/g for solids |
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Gene |
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The fundamental unit of heredity responsible for a given trait in an organism. A site on a chromosome that provides information for a certain cell function. A specific segment of DNA that contains the necessary code to make a protein or RNA molecule. |
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Genotype |
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Genetic makeup of an organism. The genotype is ultimately responsible for an organism's phenotype, or expressed characteristics. |
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Phenotype |
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The observable characteristics of an organism produced by the interaction between its genetic potential (genotype) and the environment. |
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Chromosome |
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The tightly coiled bodies in cells that are the primary site of genes. |
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Nucleotide |
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The basic structural unit of DNA and RNA; each nucleotide consists of a phosphate, a sugar (ribose in RNA, deoxyribose in DNA), and a nitrogeneous base such as adenine, guanine, cytosine, thymine (DNA), or uracil (RNA). |
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Semiconservatism |
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In DNA replication, the synthesis of paired daughter strands, each retaining a parent strand template. DNA copies itself just before cellular division by this process. Semiconservatism replication means that each "old" DNA strand is the template upon which each "new" strand is synthesized. |
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mRNA |
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Carries the message of DNA to the ribosome. Contains *codons*: a sequence of base pairs that code for an amino acid. Eg: AUG - you use the codon to determine the name of the amino acid. |
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tRNA |
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Adapter molecule to transfer RNA. Adapts the genetic code based on the base sequence of mRNA anticodon. Eg: AUG-UAC brings the amino acid from the cytoplasm into the ribosome. |
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rRNA |
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Not clear what it does but it is thought to synthesize ribosomes. A single-stranded transcript that is a copy of part of the DNA template. |
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DNA Polymerase |
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Adds new nucleotides to a new strand of DNA modeled after an old strand of DNA. It is also for proofreading. Note that the nucleotides are not stable because DNA ligase has not been applied yet. |
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RNA Polymerase |
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Enzyme process that translates the code of DNA to RNA |
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DNA Ligase |
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seals in nucleotides (composed of a phosphate, nitrogenous base, and a sugar) after DNA Ligase is applied the nucleotides are stable |
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DNA Gyrase |
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located in Bacteria cells only. "supercoiling" DNA |
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Molecular Genetics |
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When a message of DNA is turned into a protein. "Gene Expression" |
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DNA Replication |
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The semiconservative mechanisms that ensure precise duplication of the parent DNA strands. DNA fingerprinting. |
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Transcription |
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mRNA synthesis: the procss to which a strand of RNA is produced against a DNA template. |
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Translation |
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Protein synthesis; the process of decoding the messenger RNA code into a polypeptide. |
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DNA ---> DNA |
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Replication |
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DNA ---> RNA |
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Transcription |
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RNA --- > Proteins |
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Translation. Proteins are the functional products of DNA because they carry out the instructions of DNA. |
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Mutation |
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A Change in DNA sequence. Mutations can benefit the microbe, but in most cases it does not. |
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Frameshift mutation |
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Addition or deletion of one or more bases CGC GGT --- > CGC |
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Base Substitution or Point Mutation |
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One base is substituted for another base. CCT --- > CTT |
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Induced Mutations |
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Happen all the time. They result from exposure to known mutagens: 1. Chemicals 2. Radiation |
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Spontaneous mutations |
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Spontaneous mutations are very rare. Change in DNA arising from errors in replication, when there is an absence of an environmental agent. |
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Gene Expression: What are 3 ways to express a gene? |
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Constitutive Induction Repression |
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Constitutive |
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Set of genes that's constantly on |
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Induction |
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Set of genes that's turned on, so this means they are initially off. These are genes we don't need. EG: Cancer cells |
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Lactose Operon -- Induction |
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Turned on in the absence of glucose, does not make lactose, makes 3 enzymes that convert lactose to glucose. The repressor -- allosteric protein -- is what is causing the operon to be off. The lactose binds to the allosteric site causing the active site to change shape. So then the repressor comes off, allowing RNA polymerase to bind to the promoter and initiate transcription. |
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Secondary Messengers |
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Secondary messengers tell the lactose operon whether or not it is needed. |
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Tryptophan Operon - Repression |
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Makes tryptophan. When the conc. of tryptophan meets the max. threshold, Tryptophan binds to the allosteric site acting as a coil(?) repressor. The repressor protein changes shape and the conc of trypt goes down and the operon is turned off. |
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Types of Gene Transfer -- All-natural mutations |
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Conjugation Transduction Transformation |
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Conjugation -- Gene transfer between 2 organisms by a pilus |
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A pilus can only be formed by the F-plasmid (F+) which contains genetic information. F stands for fertility. An F+ donor cell and a F-recipient (F-) both contain a host cell genome. The F plasmid (F+) cell makes a copy of itself by DNA polymerase, ligase, helicase, gyrase, and semiconservatism which moves it over to the F-cell. When the F-plasmid moves over then it is no longer a recipient cell but a donor cell (F plasmid) survival of the species? |
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Transduction -- Gene Transfer between bacteria cells through a bacteriophage |
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The virus, containing nucleic acid, enters cell A's genome and leaves with genes from cell A and enters into cell B. Cell A is destroyed. So the virus now contains nucleic acid and cell A's genes. When it goes into cell B it leaves behind part of c ell |
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Tranformation |
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The uptake of naked DNA by another bacterial cell. In a host cell genome, inside of a bacteria cell, DNA comes out in fragments from the cell bursting. To make the cell burst it has to go under change in water availability (hypotonic) or temperature, or PH. This is how the DNA comes out of the cell. Then another bacteria cell can bring in one of the DNA fragments. |
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Classical Biotechnology |
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Use living organisms to produce useful products to solve a problem. "invivo" testing - using a rat or some type of animal |
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Modern Biotechnology |
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Use biological molecules to produce useful products or solve a problem. "Invitro" testing -- using a test tube with bacteria cells or human cells. |
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Genetic Engineering |
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More specific terms of biotechnology. Manipulate the genes of an organism. Recombinant DNA technology. |
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Recombinant DNA technology |
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Cutting and combining genes from 2 different organism. It involves modification of an organism's genome. Used to create genetically modified food that may have increased nutritional properties, be easier to grow, or may even vaccinate the person eating it against a host of diseases. |
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Tools of the Trade for Recombinant DNA technology |
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1. Restriction Enzymes -- enzymes that recognize and cut DNA sequences. They learn things from bacteria cells. 2. Plasmids. Genest that can make bacteria resistant to antibiotics and can express genes from foreign organisms. 1. F-plasmid. 2. R-Plasmid, R-Resistance. Carry antibiotic resistance and are self-replicating. 3. DNA Ligase - seals in the gene. |
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Transposons |
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Act Like Plasmids, mobile genetic elements -- "jumping Genes". They can jump from one plasmid or host cell genome in a DNA unit to the other and replicate itself. This causes instability in the cell. |
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Contributions to Biotechnology |
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1. Gene Library -cloned DNA stored in yeast, viruses, or bacteria. 2. Polymerase Chain Reaction (PCR)-amplify DNA. 3.DNA Fingerprinting - analyze DNA/associated with replication, forensics, paternity testing, diagnostics 4. Medicines 5. Vaccines 6. Human Genome Project |
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Examples of Aerotolerant bacteria |
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Lactobaccillus - Yoghurt Acidophilus - Makes lactic acid to keep urogenital tract low PH. Both of these are good bacteria cells that keep you from getting a yeast infection. |
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Aseptic Technique |
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Methods of handling microbial cultures, and patient specimens in a way that prevents infection of the handler and others who may be exposed. |
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Operon |
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Cluster of genes that regulate metabolism by controlling mRNA production. 1. Promoter - where RNA polymerase binds to start transcription. DNA sequence. 2. Operator -- a repressor binds to block transcription. On/Off switch. DNA sequence 3. Structural Genes -- encode for a set of enzymes. |
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Bacteriostatic |
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Any process or agent that inhibits bacterial growth |
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Bactericidal |
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Killing bacteria cells |
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Asepsis |
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A safe level free of viable pathogenic microorganisms but is not sterile. You still may see microbial growth including endospores. |
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Sterilization |
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Any process that completely removes or destroys all viable microorganisms, including viruses, from an object or habitat. Endospores are the indicators if something is sterile or aseptic. All sterile techniques are bactericidal. |
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Moist Heat |
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Heat can be under pressure or not. Heat that is under pressure is steam and is sterile, killing endospores. Example is a Crock Pot. Heat that is not under pressure is boiling. Boiling is a bactericidal, aseptic technique -- does not kill endospores. |
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Dry Heat |
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Sterile. Example: when we flame the loop in lab. |
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Cold Heat |
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bacteristatic and aseptic |
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Mesophiles |
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Grow at intermediate temperatures, warm loving. 37 degrees C. Eg: staph. aureus. |
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Psychrophiles |
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Cold-loving bacteria, thrive at low temperatures (0-20 degrees C.) |
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Thermophiles |
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Hot loving, 60 degrees C. or higher. Endospore forming bacteria. Obligate anaerobes. |
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PH affecting microbial growth |
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Bactericidal and aseptic, because endospores can survive in too basic or too acidic condition. • Acidophile --likes pH3 • Neutrophle -- likes pH7(our bodies) • Alkalophile -- likes pH9 |
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Water availability affecting Microbial Growth |
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Cells are drying. Cells like an isotonic environment. Cells are bacteristatic but over time it becomes bactericidal and aseptic. It takes a while to kill the cells because of the water. |
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Halophile |
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likes a hypertonic environment (salt-loving) Eg: staph aureus |
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Radiation affecting Microbial Growth |
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UV Radiation and Gamma Radiation |
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Explain UV Radiation |
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Starts bactericidal, but over time it becomes sterile. UV rays don't penetrate through clothes or plastic. Becomes sterile by Thymine Dimers -- fuse 2 thymine together, this blocks DNA Polymerase from reading it so the UV light can now destroy the cell. |
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Explain Gamma Radiation |
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Sterile Technique. Can go through clothes, plastic. They form free radicals. |
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Filtration. |
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Sterile technique that doesn't interfere with any cellular structures or processes and is mechanically removing bacterial cells from a solution. Once a solution goes through the filter it becomes sterile. |
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Size of Filter Unit Pores |
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.01um for bacteria and viruses .10um for bacteria and viruses .22um for bacteria but not viruses .45um for some bacteria but mainly dirt |
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Chemical Factors of Microbial Growth |
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Interfere with proteins and the plasma membrane. (mode of action) •Alcohol •Halogens |
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Describe the alcohol factor of microbial growth |
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bacteriostatic and aseptic. 70% alcohol is more effective than 100% alcohol because the water content in the 70% helps it be absorbed faster. |
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Bleach |
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Halogen that is aseptic and bactericidal. If it is used properly it is a sterile technique that interferes with DNA. |
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Iodine |
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Halogen that is bactericidal and aseptic |
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Phenols/Phenotics |
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Halogens that are bactericidal and aseptic. Eg. LIsterine/Lysol/Triclosan |
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Surfactants |
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Halogens that are bacteristatic and aseptic. Eg. Hand Soap |
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Heavy Metals |
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Halogens that are antiseptic, bactericidal, and antimicrobial (nickel, silver, mercury) |
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What are the physical factors of microbial growth? |
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Temperature, pH, water availability, radiation |
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How is DNA, mRNA, and fRNA made? |
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DNA -- 5' - 3' -- 3' - 5' mRNA 5' - 3', so the 3' - 5' from DNA is what we have to use because it is the opposite. tRNA -- 3' - 5' because it is the opposite of mRNA |
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Example of Facultative Anaerobe? |
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E. Coli and Staph. Aureus |
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Example of Microaerophile? |
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Heliobacter Pylori |
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Example of Obligate Aerobe? |
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Bordetella Pertussis |
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Where does transcription occur in Prokaryotic and Eukaryotic Cells? |
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Transcription of DNA to RNA occurs in the nucleus of Eukaryotic cells and the nucleoid of Prokaryotic cells. |
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Where does Translation occur in Prokaryotic and Eukaryotic Cells? |
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In the ribosomes. |
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Direct bacteria cell counts |
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Using the eye to see bacteria. Also uses an instrument like a coulter counter. As cells pass through this device, they trigger an electronic sensor that tallies their numbers. |
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Indirect bacteria cell counts |
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1. Metabolism - looking at by-products. Co2 and ATP are measured. 2. Dry weight - wet solution 3. Turbidity - cloudiness in the test tube. |
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Describe a mass spectrophotometer |
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Used to measure indirect bacteria cells turbidity. You place a testtube in a slot where light is coming through. The light bounces off anything solid. We get the Optimal Density from this. |
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Inaccuracy of coulter counter and spectrophotometer |
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There is no distinction between dead and live cells |
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Name examples of Obligate Anaerobes |
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Clostridium Perfringens -- gangrene Clostridium tetini -- tetanus Clostridium botulinum -- botulism |
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What is the AMPr gene? |
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Ampicillin resistance gene |
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Recombination in bacteria depends in part on the fact that bacteria contains... |
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Extrachromosomal DNA - Plasmids -- interchanging genes |
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What is a Functional Product? |
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Fu |
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What are the essential elements? |
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Carbon, Hydrogen, Oxygen, Phosphorus, Nitrogen, Sulfur, Iron. All living things require these elements. |
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Growth Factors -- Organic Nutrients |
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Only required for fastidious organisms (organisms that require special nutritional or environmental conditions for growth). Eg: nisseria gonorrhea -- needs amino acids. 1.Amino acids. 2. Purines. 3. Pyrimidines. |
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What is the advantage of using Agar? |
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Agar is a polysaccharide. It is heat stable meaning it can be heated up and it still works. Nutrients have to be placed in the agar. |
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What are the 2 categories of Agar? |
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• Complex media - used in lab. We don't know or care the exact chemical composition. • Chemically-defined agar (more expensive, unnecessary unless you're doing research, looking for particular stuff to grow or isolate) |
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Trace Elements, Inorganic? |
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Micronutrients (Zinc, copper, Magnesium) that occur in small amounts and are involved in enzyme function and maintenance of protein structure. They are non-fastidious organisms (meaning they don't require special nutritional or environmental conditions for growth. |
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Repression |
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Set of genes that are turned off. We need these genes, they are always on. They are controlled by a repressor. The repressor is not bound to the operator (so it is on) so we have to make the repressor bind to the operator to turn it off. |