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Viruses Components
- Icosahedral
- Helical
- Complex
- Enveloped
- Naked
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What is a Icosahedral?
- 3-D, 20-sided figure with 12 evenly spaced corners
- Nucleic acid is packed into he center, forming a nucleocapsid
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What is a Helical
- Have rod-shaped capsomers that bond together to form a series of hollow discs resembling a bracelet
- Discs link with other disc to form a continuous helix into which the nucleic acid strand is coiled
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What is Complex virus?
- May have have multiple types of proteins and take shapes that are not symmetrical
- Bacteriophage
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What does Enveloped mean?
- Viruses that posses an additional covering external to the capsid that is usually a modified piece of the host's cell membrane
- Envelope usually has special receptor spikes inserted into it
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What is a Naked virus?
Viruses that consist of only a nucleocapsid
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How does - sense RNA make proteins?
- RNA --> + RNA --> Proteins or - RNA
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How does + sense RNA make protein?
- + RNA --> Protein
- + RNA --> - RNA --> + RNA
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How does retrovirus make protein?
RNA --> DNA --> Protein
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What is the order of the Viral Life Cycle?
- Adsorption
- Penetration
- Uncoating
- Synthesis
- Assembly
- Release
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Viral Life Cycle
Adsorption
- Virus attaches to its susceptible host cell by specific binding of its spikes of cell receptors
- Limited host Range
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Viral Life Cycle
Penetration
- Virus is engulfed into a vesicle
- Endocytosis: Entire virus engulfed by the cell and enclosed in a vacuole or vesicle
- Fusion: Viral envelope directly fuses with the host cell membrane
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Viral Life Cycle
Synthesis
- Replication and protein production
- Under the control of viral genes, the cell synthesizes the basic components of new viruses: RNA, molecules, capsomers & spikes
- DNA enters nucleus --> Transcribed into RNA --> Translation into viral proteins --> New DNA synthesized using host nucleotides
- RNA viruses replicated and assembled in the Cytoplasm
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Viral Life Cycle
Assembly
Viral spike proteins are inserted into the cell membrane for the viral envelope; nucleocapsid is formed from RNA and capsomers
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Viral Life Cycle
Release
- Enveloped Viruses bud off of the membrane, carrying away an envelope with the spikes
- Noneveloped and Complex viruses released when they lyses or ruptures
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What is Cytopathic Effects?
- Virus-induced damage to the cell that alters its microscopic appearance
- Used to identify virus infections
- Inclusion bodies: Compacted masses of viruses or damaged cell organelles in the nucleus and cytoplasm
- Syncytia: Fusion of multiple host cells into single large cells containing multiple nuclei
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What is Viral Transformation?
- When viruses enter their host cell and permanently alter its genetic material, leading to cancer (oncoviruses)
- Cells Lose Contact Inhibition:
- Immortalization
- Causes cells to have
- -Increases rate of growth
- -Alterations in chromosomes
- -Changes in the cell's surface molecules
- -Capacity to divide for an indefinite period
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What is a bacteriophages?
Viruses that infect bacteria
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In the Lytic Cycle, what happens in the Eclipse Phase?
- Phage is developing but is not yet infectious
- -Adsorption
- -Penetration
- -Duplication of phage components; Replication of virus genetic material
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In the Lytic Cycle, what happens in the Virion Phase?
- virus matures and is capable of infecting a host
- -Assembly of new virions
- -Maturation
- -lysis of weakened cell
- -Release of viruses
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What happens in the Lysogenic Cycle:
- Host chromosome carries bacteriophage DNA
- DNA phages undergo adsorption and penetration into the bacterial host and then enter an inactive prophage stage, during which it is inserted into the bacterial chromosome
- Viral DNA retained by the bacterial cell and copied during its normal cell division so that the cell's progeny will also have the temperate phage DNA
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What is a Prion?
Infectious protein that contains no DNA or RNA
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How does Prion cause diseases?
- An abnormal prion protein is acquired via ingestion or mutation
- Infectious protein induces mis-folding or host protein
- -Mis-folded protein can change other proteins
- -Protein plaques and vacuoles harm neurological function
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What are some examples of Prion Diseases?
- Mad Cow Disease (Bovine Spongiform Encephalopathy [BSE])
- Cruetzfeld-Jakob Disease
- Variant Creutzfeldt-Jakob
- Kuru
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Define Genome
Sum total of genetic material of a cell
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Define Chromosome
Discrete cellular structure composed of neatly packaged DNA molecule
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Define Gene
Certain segment of DNA that contains necessary code to make a protein or RNA
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Define Phenotype
Traits created by the expression of the genotype
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Define Genotype
An organism's distinctive genetic makeup from the sum of all types of genes
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What is a Hydrogen Bond?
- Join Nitrogenous bases
- Weak bonds that are easily broken, allowing the molecule to be unzipped into its complementary strands
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What is a Covalent Bonds?
Nucleotides bond to form a sugar-phosphate linkage that becomes the backbone of each strand
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What is a Nucleotide?
- Basic unit of DNA structure
- composed of phosphate, deoxyribose sugar and a nitrogenous base
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What are the parts of Nitrogenous Bases?
- Purines and Pyrimidines
- Attach by covalent bonds at the 1' position of the sugar
- Spanned the center of the molecule & pair w/ complementary bases from other grands
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Which enzymes are involved in DNA Replication?
- Helicase: Unzip the DNA helix
- Primase: Synthesize an RNA primer
- DNA Polymerase III: Add base to new DNA chain; proofread chain for mastakes
- DNA Polymerase I: Remove primer, close gaps, repair mismatches
- Ligase: Final binding of nicks in DNA during synthesis and repair
- Gyrase: Supercoil
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Origin of Replication and How it Starts
- Origins of Replication: Short sequence of rich in A and T held together by only two H-bonds
- Helicases bind to the DNA at the origin
- -Untwist the helix
- -Break the H-bonds
- -Results in two separate strands
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Leading Strand vs. Lagging Strand
- Leading Strand: Synthesized as a continuous, complete strand in 5' to 3' direction
- Lagging Strand: cannot be synthesized continuously due to the opposite orientation (3' to 5')
- -Polymerase adds nucleotides a few at a time in the direction away from the fork (5' to 3')
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What are Okazaki Fragments?
- Fragments of DNA on the Lagging Strand
- Few nucleotides added at a time in the 5' to 3' direction
- Fork opens & next segment is synthesized backward to point of previous segment
- Fragments attached to the growing end of the lagging strand by DNA Ligase
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What is Transcription?
Master code of DNA is used to synthesize an RNA molecule
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RNA Structure, Nucleotides and Base Pairing
- Single-stranded molecule that exists in helical form
- Contains Uracil instead of Thymine
- Sugar in RNA backbone (alternating with phosphate) is Ribose (not Deoxyribose)
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Enzymes in RNA
- RNA Polymerase: Unwinds the DNA at the promotor
- -Moves along the strand adding complementary nucleotides as dictated by the DNA template, forming the single-stranded mRNA
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Promoter, Termination Sequence in Transcription
- Promotor: Region of two sequences of DNA just prior to the beginning of the gene to be transcribed
- Recognized by the RNA polymerase
- Termination Sequence: RNA Polymerase recognizes a code that signals the separation and release of the mRNA strand
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What is the Direction of Transcription?
- RNA Polymerase runs along the template strand in the 3' to 5' direction
- Elongation proceeds in the 5' to 3' direction
- The mRNA reads in the 5' to 3' direction
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What are the Components of Translation?
- Ribosomes: Located in the cytoplasm
- Subunits specifically adapted to assemble & form sites to hold the mRNA & tRNA
- Recognizes these molecules and stabilizes reactions between them
- tRNA: Brings amino acids to ribosome during translation
- Anticodon that designates the specificity of the tRNA & complements mRNA's codons
- mRNA: Sequence of amino acids in protein
- Carries the DNA master code to the ribosome and then translated
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What are the steps of translation?
- Initiation
- Elongation
- Termination
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What happens in Initiation?
- Small subunit binds to the 5' end of mRNA and large subunit supplies enzymes for making peptide bonds on the protein
- Begins to scan the mRNA by moving in the 5' to 3' direction along the mRNA
- The first codon is the START codon (AUG)
- mRNA message in place on the ribosome, tRNA enter with their amino acids
- -Complementary tRNA (anticodon) meets with the mRNA code
- -Guided by the two sites on the large subunit called the P site & A site
- -E site is where the tRNA's are released
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What happens in Elongation during Translation?
- Ribosome shifts its reading frame to the right along the mRNA from one codon to the next
- -Brings the next codon into place on the ribosome and makes a space for the next tRNA to enter the A site
- -Peptide bond is formed between the amino acids on the adjacent tRNA's
- -Polypeptide grows in length
- Translocation: Enzyme-directed shifting of the ribosome to the right along mRNA strand, which causes bland tRNA to be discharged from the ribosome at the E site
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What happens in Termination during Translation?
- Brought about by the presence of a Termination Codon - UAA, UAG, & UGA
- -No corresponding tRNA (or amino acid)
- -Nonsense Codons
- Special enzyme breaks the bond between the final tRNA and the finished polypeptide chain
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Define Redundancy
A particular amino acid can be coded for by more than a single codon
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Define Wobble
- Only the first two nucleotides are required to encode the correct amino acid
- -Permits some variation or mutation without altering the message
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What happens in Prokaryote Transcription & Translation?
- Translation of mRNA starts while transcription is still occurring
- Increases efficiency
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What happens in Eukaryote Transcription/Translation?
- Start codon is also AUG but may code for an amino acid (methionine)
- mRNAs code for just one protein
- DNA is in nucleus so transcription and translation cannot be simultaneous
- mRNA must pass through pores in the nuclear membrane and be carried to ribosome's in the cytoplasm
- Introns: Sequences of bases that do not code for protein
- Exons: Coding regions that will be translated into protein (Express)
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In gene regulation, what is prokaryotic operons?
Coordinated set of genes, all of which are regulated as a single unit
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What is Lac operon? (Inducible)
- Operon is turned on by the substrate of the enzyme for which the structural genes code
- -Normally off; Adding Lactose turn it on
- Operon Off: in the absence of lactose, a repressor protein attaches to the operator of the operon
- -Locks the operator & prevents any transcription of the structure gense downstream
- -Supression of transcription prevents the unneccessary synthesis of enzymes for processing lactose
- Operson On: Upon entering the cell, lactose (substrate) becomes a gentic inducer by attaching to the repressor, which loses its grip and falls away
- The RNA polymerase is free to bind to the promotor and initiate transcription
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What is Arg operon? (Repressible)
- Contain genes coding for anabolic enzymes
- Several genes in a series are turned off by the product synthesized by the enzyme
- -Normally on; Turned off when the nutrient is not longer required
- Operon On: A repression operon remains on when its nutrient products are in great demand by the cell
- -The repressor is unable to bind to the operator at low nutrient levels
- Operon Off: The operon is repressed when
- -Arginine builds up & serves as a corepressor, activating the the repressor (negative feedback)
- -Repressor complex affixes to the operator and blocks the RNA polymerase and further transcription of genes for arginine synthesis
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Define mutant
Microorganisms that bears a mutation (Phenotypic changes due to the changes in the genotype)
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Define Wild Type
Microorganism that exhibits a natural, non-mutated characteristic
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Spontaneous vs. Induced
- Spontaneous: Random change in the DNA arising from errors in replication that occur randomly
- Induced: Result from exposure to known mutagens (physical or chemical agents that interact with DNA in a disruptive manner)
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What are the types of mutation and the results to protein function?
- Point Mutation: Involve addition, deletion, or substation of single bases
- -May or may not change protein
- Missense Mutation: Any change in code that leads to placement of a different amino acid
- -Can create a faulty, nonfunctional protein
- -Can produce a protein that functions in a different manner
- -Can cause no significant alteration in protein function
- Nonsense Mutation: Changes a normal codon into a stop codon
- Stops the production of the protein wherever it occurs
- Almost always results in a nonfunctional protein
- Silent Mutation: Alters a base but does not change the amino acid
- -Has no effect on the protein function
- Back-Mutation: Gene that has undergone mutation reverses to its original base composition
- Frameshift Mutation: Mutations that occur when one or more bases are inserted into or deleted from a newly synthesized DNA strand
- -Changes the reading frame of the mRNA
- -Nearly always result in a nonfunctional protein
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Mutation Repair
- Enzymatic Enzymes: Special for finding and fixing such mutations; Most common way
- DNA that has been damaged by UV radiation
- -Restored by Photoactivation or Light Repair
- --DNA photolyase- Light sensitive enzyme
- Excision Repair
- -Excise mutations by a series of enzymes
- -Remove incorrect bases and add correct one
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Define Recombination
When one bacterium donates DNA to another bacterium
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In recombination, what happens in Conjugation?
- Requires the attachment of two related species and the formation of a bridge that can transport DNA
- -Physical Conjugation: The pilus of donor cell attaches to receptor on recipient cell and retracts to draw the two cells together
- --An opening or pore forms between the cell walls, creating a bridge to transmit genetic material
- -F Factor Transfer: F Factor (plasmid) copies and then transferred though a bridge
- -H factor Transfer: F factor integrated into chromosome and then duplicated and transmitted in part to a recipient cell, where it is integrated into the chromosome
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In recombination, what happens in Transformation?
- Entails the transfer of naked DNA and requires no special vehicle
- -Recipient cell takes up donor DNA
- -Donor DNA align with complementary bases
- -Recombination occurs between donor DNA and recipient DNA
- -Griffith's Experiment
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In recombination, what happen in Transduction?
- DNA transfer mediated though action of a bacterial virus (bacteriophage)
- Generalized:
- -Random fragments of disintegranting host DNA are taken up the the phage during assembly
- -Any gene from the bacterium can be transmitted
- -Lytic Infection: Lyses the cell to infect others
- --Releases the mature phases, including the genetically altered one
- Specialized:
- -Highly specific part of host genome is regularly incorporated into the virus
- -Lysogenic Infection: Integrated in bacterial chromosome
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What happens in Transposons?
- Transposable elements
- Shift from one part of the genome to another
- -From one chromosomal site to another
- -From a chromosome to a plasmid
- -From a plasmid to a chromosome
- Contain DNA that codes for the enzymes needed to remove and reintegrate the transposon at another site in the genome
- Overall Effect- Scrambles the genetic language
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How is heat use to manipulate DNA?
- Heating to 90oC separates DNA strands exposing nucleotides
- Cooling to allow annealing
- Complementary strands from different organisms hybridize
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How does Restriction Enonucleases manipulate DNA?
- Enzymes that can clip strands of DNA crosswise at selected positions
- Can recognize and clip at Palindromes (Identical sequences of DNA when read from 5' to 3' direction on one strand and the 5' to 3' direction on the other strand)
- Used to cut DNA in to smaller pieces for study or remove & insert sequences
- Can make a blunt cut or a "sticky end"
- Restriction Fragments: The pieces of DNA produced
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What is cDNA?
- Copies made from messenger, transfer, ribosomal and other forms of RNA
- Synthesize eukaryotic genes from mRNA transcripts
- Synthesized gene will be free of the intervening introns
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What is Electrophoresis?
Produces a readable pattern of DNA fragments
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How does Electrophoresis works?
- Samples placed in wells in a soft agar gel and subjected to an electrical current
- Phosphate groups in DNA give the molecule an overall negative charge
- -Causes DNA to move toward the positive pole in the gel
- Rate of movement based on size of fragments
- -Larger fragment move slowly and remain near the top of the gel
- -Smaller fragments move faster and positioned farther from the wells
- Positions of DNA fragments determined by staining the DNA fragments in the gel
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What information can you get with Electrophoresis?
- Characterize DNA fragments and compare the degree of genetic similarities among samples
- Genetic Fingerprint
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PFGE (Pulsified-Field Gel Electrophoresis)
- Macrorestriction analysis
- Pathogens isolated from a patient and their DNA is harvested
- -DNA is cut up with restriction enzymes to get a few very large pieces of DNA
- -DNA separated using the pulsed-field method of gel electrophoresis
- --Constantly changes the direction of (pulsing) the electrical field
- --Allows effective separation of the large pieces
- Fragments of different lengths seen as dark bands after special stain on gel
- Patterns different t/t enzyme cut in different places on genome where small DNA changes exist, correspond to different strain types
- DNA Fingerprint
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Define Nucleic Acid Hypridization
Two different nucleic acids hybridize by uniting at their complementary regions
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Define Gene Probes
- Specifically formulated oligonucleotide tracers
- Consists of a short stretch of DNA or a known sequence that will base-pair with a stretch of DNA with a complementary sequence (if in the test sample)
- Carry reporter molecules so areas of hybridization can be visualized
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What is a Southern Blot?
- Type of Hybridization
- DNA fragments are first separated by electrophoresis and then denatured and transferred to a special filter
- DNA probe incubated with the sample
- -Wherever this probe encounters the segment of which it is complementary, it will attach and form a hybrid
- Development of hybridization pattern will show up as one or more bands
- Sensitive and specific way to isolate fragments from a complex mixture and to find specific gens sequences on DNA
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What is FISH (Fluorescent in Situ Hybridization)?
- Probes applied to intact cells
- Observed microscopically for the presence and location of specific genetic marker sequences
- Effective way to locate genes on chromosomes
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What is DNA sequencing?
- Determines the actual order and types of bases in DNA
- Sanger Method: Most common technique
- Limitations:
- -Sanger technique is old and expensive (reaching the end of its useful life)
- -May not be specific - None-specific binding of the primer to the DNA
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What is PCR (Polymerase Chain Reaction)?
- Rapidly increases the amount of DNA in a sample
- How it is done
- -Denaturation: Heat to separate into two strands
- -Priming: Primers added in a concentration that favors binding to the complementary strand of test DNA
- --Prepares the two strands (amplicons) for synthesis
- -Extension: DNA Polymerase and nucleotides are added to primer
- -Cycling: 20-30 cycles to amplify DNA
- Results: Amplified DNA to be anayzed
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What is Recombinant DNA Technology?
- Deliberately removes genetic material from 1 organisms & combines it with that of a different one
- -Forms genetics closes
- --Genes is selected
- --Excise gene
- --Isolate gene
- --Insert gene into a vector
- --Vector insets DNA into a cloning host
- What are the results?
- -Mass produce substances that are difficult to synthesize by the usual industrial methods
- --Hormone, enzymes, vaccines
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What is a vector?
- Plasmid or virus that a gene is inserted into
- Inserts the DNA into a Cloning Host
- Characteristics
- -Capable of carrying a significant piece of the donor DNA
- -Must be readily accepted by the cloning host
- -Must have a promotor in front of the cloned gene
- -Origin of Replication needed somewhere so that it will be replicated by the DNA polymerase of the cloning host
- -Must accept DNA of the desired size
- -Contain a gene that confers drug resistant to their cloning host
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What is a host?
- Bacterium or yeast that can replicate the gene & translate it into the protein product
- Characteristics
- -Rapid turnover, fast growth rate
- -Can be grown in large quantities using ordinary culture methods
- -Nonpathogenic
- -Genome that is well delineated
- -Capable of accepting plasmid or bacteriophage vectors
- -Maintains foreign gene though multiple generations
- -Will secrete a high yield of proteins from expressed foreign genes
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What is Gene Therapy?
Correcting or repairing a faulty gene in humans suffering from a fatal or debilitating disease.
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How does Antisense RNA work in gene therapy?
- Bases are complementary to the sense strand of mRNA in the area surrounding the initiation
- -When it binds to the mRNA, the dsRNA is inaccessible to the ribosome
- -Translation cannot occur (stop bad proteins)
- Genetic medicine
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How does Antisense DNA work in gene therapy?
- When delivered into the cytoplasm and nucleus, it binds to specific sites on any mRNAs that are the targets of therapy
- -Antisense Therapy: Treat Cytomegalovirus Retinitus
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Define Disinfection
Destroys most microbial life, reducing contamination on inanimate surfaces
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Define Sterilization
Destruction of all microbial life
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Define Antisepsis
Same as disinfection except a living surface is involved
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Define Sanitation
Any cleansing technique that mechanically removes microorganisms to reduce contamination to safe levels
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What are the 2 most resistant microbes?
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Killing curves
- Factors that influence the rate at which microbes are killed by antimicrobial agents
- Length of Exposure to Agent
- Effect of the Microbial Load
- Relative Resistance of Spores vs. Vegetative Forms
- Action of the Agent (Microbicidal or Microbiastatic
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What are the mechanisms to kill bacteria?
- Change Membrane permeability
- -Lose selective permeability so can't prevent the loss of vital molecules or stop the entry of damaging chemicals
- Damage the Cell Wall
- -Block its synthesis
- -Digest it
- -Break down its surface
- -The cell becomes fragile and is lysed easily
- Damage Protein
- -denature (disrupt) proteins
- Damage Nucleotides
- -Binding to ribosomes to stop translation
- -Bind irreversibly to DNA preventing transcription and translation
- -Mutagenic agents
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How is head use to control bacteria?
- Moist Heat: occurs in the form of hot water, boiling water, or steam
- -Ranges from 60 to 135 Celsius
- Dry Heat: Denotes air with a low moisture content that has been heated by a flame or electric heating coil
- -Ranges from 160 to several thousand degrees of Celsius
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Autoclaving
- Steam under pressure
- Pressure raises the temperature of steam
- Most efficient pressure-temperature combination for sterilization: 15 psi which yiels 121 degree celsius for 15 minutes
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Pasteurization
- Heat is applies to liquids to kill potential agents of infection and spoilage *disinfect)
- Maintains the liquids' flavor and food values
- Does not kill endospores or thermoduric microbes
- Does not sterlize
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Boiling
- Disinfection
- Expose material to boilng water for 30 minutes
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Baking in Oven
- Used for heat-resistant items that do not sterilize well with moist head
- -Glass, metallic instruments; powders, oils
- Sterilization
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Incineration
- Ignites and reduces microbes to ashes and gas
- Sterlizes
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What does Ionizing Radiation do?
- Cells' molecules absorb some of the available energy
- Radiation ejects orbital electrons form an atom, causing ions to form
- Gamma Rays, X Rays & Cathode Rays
- Cold sterilization
- Uses
- -Food products
- -Medical products
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What does UV (Nonionizing Radiation) Do?
- Excites atoms by raising them to a higher energy state, but it does not ionize them
- Atomic excitation leads to formation of abnormal bonds within molecules and is a source of mutations
- Not as penetrating as ionizing radiation
- doesn't kill bacterial spores
- Uses
- -Disinfect: Hospital rooms, operating rooms, schools, food prep areas, dental offices
- -Treat drinking water or purify liquids
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Filtration
- Mechanical technique for removing microbes
- Effective for removing microbes from air and liquids
- Fluid strained through a filter with openings large enough for fluid but too small for microorganisms
- Filters are usually thin membranes of cellulose acetate materials
- Pore size can be standardized
- Prepare liquids that can't withstand heat
- Can't decontaminate beverages without altering their flavor
- Water purification
- Removing airborne contaminants (HEPA filters)
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Examples of Chemical
- Halogen - betadine; Bleach
- Phenol- Listerine; Lysol
- Chlorhexadine- Avagard
- Alcohols- Hand sanitizer
- Hydrogen Peroxide
- Detergents- Soaps
- Heavy metal compounds- Mercury; Silver; Silver nitrate
- Aldehydes- Glutaraldhyde; Formaldehyde
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What chemical can be used on people?
- Halogen- Yes
- Phenol- No
- Chlorohexadine- Yes (had scrubbing, preparing skin for surgery, neonatal wash)
- Alcohols- Yes
- Hydrogen peroxide- Yes
- Heavy metals- Yes (topical)
- Aldehydes: No
- Gases
- -Ethylyne Oxide- No
- -Hydrogen Peroxide- No
- Dyes- Yes
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Define Prophylasix
Use of a drug to prevent imminent infection of a person at risk
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Antibiotics
Substances produced by the natural metabolic processes of some microorganisms than can inhibit or destroy other microorganisms
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Define narrow Spectrum
Antimicrobial effective against a limited array of microbial types (e.g., gram-positive bacterial)
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Define Broad Spectrum
Antimicrobial effective against a wide variety of microbial types (e.g., both gram-positive and gram-negative bacteria)
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Mechanism of Action of Antimicrobial
- Inhibition of Cell Wall synthesis
- Inhibition of Nucleic Acid Structure and Function
- Inhibition of Protein Synthesis
- Interference with Cell Membrane Structure or Function
- Inhibition of Folic Acid Synthesis
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Affect Peptidoglycan
- Bind and block peptidases that cross-link the glycan molecules
- Interrupts the completion of the cell wall
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Cephalosporins vs. Penicillins
- Resistant to most beta lactamases (penicillinases)
- Cause fewer allergic reactions than penicillins
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PBP's (Penicillin Binding Proteins)
- Catalyze cross-linking of peptidoglycan chains
- Penicillin must bind to them to produce antibacterial effects
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Why so many PCNs?
- Involved in different stages of bacterial cell wall synthesis
- When one type is inhibited, it inhibits the specific stage of peptidoglycan synthesis
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Beta lactamases
Enzymes that are cable of destroying the beta-lactam right of penicillin
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Not beta lactam
Vancomycin (Important for treating MRSA)
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Affect Nucleic Acid
- List of Actions
- -Block synthesis of nucleotides
- -Inhibit replication
- -Stop transcription
- -Inhibit DNA synthesis
- Metronizadole for parasites
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Affect Protein Synthesis
- Inhibit translation by reacting with the ribosome-mRNA comlex
- Classes of Antibiotics
- Aminoglycoside
- Tetracyclines
- Oxazolidonones
- Chloramphenicol
- Macrolides
- Licosamides
- Quinolones
- Stretogramins
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Affect Cell Membrane
- Disruption in metabolism
- Polymyxin B
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Affect Folic Acid Synthesis
- Supplied to cells in high concentrations to make sure enzyme is constantly occupied with the metabolic analog rather than the true substrate
- Sulfonamides and Trimethoprim: Competitive Inhibition
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For main groups of Antifungals
- Macrolide Polyene Antibiotics
- -Bind to fungal membranes and cause loss of selective permeability
- Griseofulvin
- -Especially active Skin fungal infections
- Synthetic Azoles
- -Broad spectrum antifungal agents
- -Ketaconazole, fluconazole,...
- Flucytosine
- -Can be used to treat certain cutaneous mycoses
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Antiparastitic
- Some also antibiotics
- Metronizadole (Flagyl)
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Antiviral Problems
Selective toxicity is almost impossible to achieve because a single metabolic system is responsible for the well-being of both virus and host
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Antiviral
Three Major modes of Action
- Barring penetration of the virus into the host cell
- Blocking the transcription and translation of viral molecules
- Preventing the maturation of viral particles
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Antiviral
Interferon
- Glycoprotein produced by fibroblast and leukocytes in response to various immune stimuli
- Produced by recombinant DNA technologies
- Known therapeutic benefits:
- Reducing the time of healing and some of the complications in certain infections
- Prevent or reducing some symptoms of cold and papilomaviruses
- slowing the progress of certain cancers
- Treating a rare Cancer called hairy-cell leukemia, hepatitis C, genital warts, and Kaposi's sarcoma in AIDS patients
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Antibiotic Resistance
Mechanisms of Resistance
- Drug Inactivation
- -Due to now enzymes being synthesized
- -Beta lactamases
- Decreased Permeability
- -Activation of Drug pumps: Uptake of drug into bacterium is decreased/eliminated
- Change in Drug binding site
- -Binding sites decreased in number or affinity
- -Mec A
- Use of Alternate metabolic pathway
- -Affected metabolic pathway shut down or an alternate pathway is used
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Negative Side Effects
- Toxic
- Allergies
- Suppression of normal flora
- Superinfection
- -Complication that occurs when antimicrobial therapy destroys beneficial resident species, causing microbes that were once in small numbers begin to overgrown and cause disease
- C. difficile example
- -Oral therapy with tetracycline, clindamycin, and broad-spectrum PCN and cephalosporins is associated with antibiotic-associated colitis due to C. Difficile
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E-test
- Uses a strip to produce a zone of inhibition
- Strip contains a gradient of drug calibrated in micrograms
- MIC can be measured by observing the mark on the strip that corresponds to the edge of the zone of inhibition
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Kirby-Bauer
- Agar diffusion test that provides useful data on antimicrobial susceptibility
- Surface of a plate of special medium is spread with the test bacterium
- Small disc containing a premeasured amount of antimicrobial dispensed onto bacterial lawn
- Zone of inhibition surrounding disc measured and compared with a standard for each drug
- Profile of antimicrobial sensitivity provides data for drug selection
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MIC (Minimum Inhibitory Concentration)
- Smallest concentration (highest dilution) of drug that visibly inhibits growth
- Useful in determining the smallest effective dosage of drug & in providing a comparative index against other antimicrobials
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Reasons for Treatment Failure
- The inability of the drug to diffuse into that body compartment
- A few resistant cells in the culture that did not appear in the sensitivity test
- An infection caused by more than one pathogen, some of which are resistant to the drug
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Define True Pathogen
Capable of causing disease in healthy person with normal immune systems
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Define Opportunistic Pathogen
Cause disease when the host's defenses are compromised or when they become established in part of the body that is not natural to them
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Define Acute disease
Infections that come on rapidly, when severe but short-live effects
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Define Chronic Disease
Infection that progress and persist over a long period of time
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Define Subacute infection
Inapparent, asymptomatic
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Define Virulence
- Describes the degree of pathogenicity
- -Determined by its ability to
- --Establish itself in the holt
- --Cause damage
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Portal of entry
A route in which the microbe enters the tissues of the body
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Transplacental or Congential Infection
- Comes across the placenta and invade the fetal circulation
- Considered a vertical transmission
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ID50
- How many organisms need to make 50% of people sick
- How does it affect Virulence?
- -Microorganisms with smaller IDs have greater virulence
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