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Life 102, Week 14 Notes

by: Sydney Dingman

Life 102, Week 14 Notes Life 102

Marketplace > Colorado State University > Biology > Life 102 > Life 102 Week 14 Notes
Sydney Dingman
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This goes over the end of Chapter 19 notes and all of 20.
Attributes of Living Systems
Erik N Arthun
Class Notes
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This 8 page Class Notes was uploaded by Sydney Dingman on Thursday April 21, 2016. The Class Notes belongs to Life 102 at Colorado State University taught by Erik N Arthun in Winter 2016. Since its upload, it has received 17 views. For similar materials see Attributes of Living Systems in Biology at Colorado State University.


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Date Created: 04/21/16
LIFE 102, Week 14 Notes 4/18/16, Chapter 19 cont.  Lysogenic Cycle o The lysogenic cycle replicates the phage genome without destroying the host o The viral DNA molecule is incorporated into the host cell’s chromosome o This integrated viral DNA is known as a prophage o Every time the host divides, it copies the phage DNA and passes the copies to daughter cells o An environmental signal can trigger the virus genome to exit the bacterial chromosome and switch to the lytic mode  RNA as Viral Genetic Material o The broadest variety of RNA genomes is found in viruses that infect animals o Retroviruses use reverse transcriptase to copy their RNA genome into DNA  Reverse transcription: RNA template  DNA o HIV (human immunodeficiency virus) is the RNA retrovirus that causes AIDS:  Acquired Immunodeficiency Syndrome  HIV Reproductive Cycle o The viral DNA is integrated into the host genome is called a provirus o Unlike a prophage, a provirus remains a permanent resident of the host cell o The host’s TNA polymerase transcribes the proviral DNA into RNA molecules o The RNA molecules function both as mRNA for synthesis of viral proteins and as genomes for new virus particles released from the cell  HIV: an RNA retrovirus o RNADNAinserts in host genomeNew viral RNA, proteins virus assembly, cell lysis  Transcription, translation  Protection from Viral Infections o Viral infections cannot be treated by antibiotics o Vaccines are harmless derivative of pathogenic microbes that stimulate the immune system to mount defenses against the harmful pathogen o Vaccines can prevent certain viral illnesses o Antiviral drugs can help to treat, though not cure, viral infections  Emerging Viruses o Emerging viruses are those that suddenly become apparent  Human Immunodeficiency Virus  Ebola virus  West Nile virus o Viral diseases in a small isolated population can emerge and become global o New viral diseases can emerge when viruses spread from animals to humans o Viral strains that hump species can exchange genetic information with other viruses to which humans have no immunity  Epidemics o Recently, a general outbreak (epidemic) of a flu-like illness appeared in Mexico and the United States, caused by an influenza virus names H1N1 o Flu epidemics are caused by new strains of influenza virus to which people have little immunity  Prions: The simplest of infectious agents o Prions are slow-acting, virtually indestructible infectious proteins that cause brain diseases in mammals o Prions propagate by converting normal proteins into the defectively-folded prion version o Scrapie in sheep, mad cow disease, chronic wasting disease, and Creutzfeldt-Jakob disease in humans are all caused by prions o Prions propagate by converting normal proteins into the defectively-folded prion version 2 4/18/16, Chapter 20: Biotechnology  The manipulation of organisms or their genetic components to make useful products  Genetic engineering: the artificial manipulation of genetic information o Changing the properties of organisms o Tailor them for our benefit o Other examples: sequencing DNA from a crime scene can identify a criminal by genotype, not phenotype o Diseases normally treated by protein-replacement therapy might be treated by actually fixing the underlying mutation  Applications of Genetic Engineering o Agriculture, Medicine, Genetic Screening, Forensics, Pollution Cleanup  DNA Technology o Recombinant DNA Techniques  Isolate DNA molecules  Cut & Paste  recombine them  Transfer to organism o Genetically engineered organism  Transgenic organism  GMO  Tools used in DNA technology o Enzymes that can modify DNA o Vectors that can carry foreign DNA into a host cell and replicate there o Hosts to replicate the DNA of interest  Enzymes: Cut & paste DNA from different sources o Cloning Vector: Bacterial plasmids  Small circular DNA molecules that replicate separately from the bacterial chromosome o Cloning Host: bacterium itself  Enzymes used in DNA technology 3 o Restriction enzymes  Bacterial enzymes that cut DNA molecules at specific DNA sequences  Cut DNA in a staggered way, producing fragments with “sticky ends”  Sticky ends can be used to join DNA from different organisms by cutting both organisms’ DNA with the same restriction enzyme  All fragments will have complementary ends o DNA Ligase  Enzyme that seals the bonds between DNA fragments  Enzymes o Cut vector and gene of interest: restriction enzymes o To paste vector and gene of interest together: ligase o DNA polymerase  Used for polymerase chain reaction  1 DNA copy  billions of copies  Making copies of DNA sequences: Polymerase Chain Reaction (PCR) o Small amounts of DNA or RNA are limiting factor in some tests aimed at analyzing the genetic information o PCR rapidly-increases the amount of DNA in a sample o Can amplify a target DNA from a few copies to billions in a few hours! o Can detect a single cancer cell or a single gene from a pathogen! o Cycle:  Denaturation  Heat to 94 degrees Celsius to separate DNA into 2 strands  Cool to 50-65 degrees Celsius  Priming  Primers added, bind to complementary strand of test DNA  Prepares the 2 DNA strands for synthesis 4  Extension  72 degrees Celsius  DNA polymerase and nucleotides are added  Polymerases extended the molecule  Repeat the steps 25-40 times 4/20/16, Chapter 20 cont.  PCR o Target DNA sequence is amplified o Automated PCR machines can perform 20 cycles on ~100 samples in 1-2 hours  Enzymes (1) o mRNA  Reverse transcriptase  complementary DNA (cDNA)  no introns  cDNA from Eukaryotes o Need to get rid of intron sequences o Isolate mRNA o Reverse transcription o mRNAcDNA (free of introns)  Cloning Vectors (2) o Isolated genes are typically spliced into “cloning vectors”  Thousands are available commercially o Examples of Vectors:  Plasmids (transferred by host by transformation)  Bacteriophages (naturally inject DNA into host via transduction)  Cloning Hosts (3) o Bacteria or Yeast o A problem with expressing Eukaryotic genes in prokaryotic cells: prokaryotic cells (such as E. coli) cannot splice mRNA because their genes do not have introns  How are genes cloned? 5 o Isolate and propagate o Make a gene library o Collection of all the genes from a cell o Pick out the gene of interest  Genomic Library o DNA library from the entire genome (100,000)  cDNA library o DNA library from the expressed genes o ~5000  5000 mRNAs  Reverse Transcriptase  5000 cDNAs  How is the Gene of Interest Identified? o Use a molecule that recognizes gene/gene product (protein)  Nucleic Acid Probe”  Complementary DNA sequence that binds to gene of interest  Antibody  Protein that binds specifically to another protein  Nucleic Acid Hybridization and Probes o Two different nucleic acids can “hybridize” by uniting at their complementary regions o Nucleic acid probes:  Short stretches of ssDNA of a known sequence  Will base-pair with ssDNA containing a complementary sequence  Can detect specific nucleotide sequences in unknown samples  Probes carry reporter molecules so they can be visualized  “labeled” via fluorescence, radiation, luminescence, or enzymes o Example of Hybridization Technique:  Southern Blot  Gel Electrophoresis 6 o Used for DNA analysis o DNA samples are placed in wells in an agarose gen  Subjected to an electrical current o (-) charged DNA passes through gel toward (+) charge  Large fragments move slowest  Small fragments move fastest o DNA fragments are stained and observed for a pattern o Produces a readable pattern of DNA fragments for characterization and comparison  “Southern Blot” o Locates specific DNA sequences o DNA is run on a gel o Transfer to membrane o Add labeled probe and let it hybridize o Wash off any unbound probes o Detect bound probes using label  Using a cloned gene to transform an organism o Add regulatory sequences (promoter, etc.) o Add selection marker (resistance gene) o Transform gene construct to cell o Regenerate transformed cell into organism  Biotechnology applications o Diagnostics: diseases, pregnancy o Genetic screening o Medicine production: insulin, growth factors o Forensics: rape/murder suspects o Biofuel production: oils, plastics o Bioremediation: pollution cleanup o Crop improvement: resistance, yield  Genetic Treatments: Gene Therapy o Some diseases are due to the lack of a specific protein 7  Type I Diabetes is caused by a lack of insulin o Goal of Gene Therapy: to permanently cure a physiological dysfunction by correcting or repairing a faulty gene o Two strategies:  In vivo: naked normal DNA or cloned virus vector is introduced directly into patient tissues  Ex vivo:  Clone normal gene in virus vector  Incubate virus with tissues removed from patient  Reintroduced transfected cells into patient o Examples of uses: immunodeficiency diseases, hemophilia, cancers… o Some diseases are due to the lack of protein o Introduce good protein by introducing normal gene o Phenotype is corrected 8


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