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Genetics Viruses & Bacteria Recombinant Technology Ch. 19 & 20 Viruses Contain nucleic acid (DNA or RNA) Protein coat – called CAPSID, protein subunits called CAPSOMERES Some have ENVELOPE – lipids and proteins from host cell Lytic Infection- death of host cell Virus penetrates host cell Viral genetic material uses host enzymes to replicate and produce viral proteins Newly assembled viruses erupt from host cell (infect new cells) Lytic Infection DNA viruses Behaves as host DNA Utilizes host enzymes RNA viruses Behave as mRNA (or template) Retroviruses – reverse transcriptase creates DNA copy of RNA to behave as host DNA Lysogenic Infection- does not destroy host Viral DNA temporarily incorporated into host DNA as a PROVIRUS Provirus replicated along with host DNA Remains inactive until environmental triggers Activation leads to lytic cycle Bacteria Single, circular DNA “Naked” chromosome – no associated proteins (like histones) Reproduction: binary fission Plasmids – short, circular DNA (beneficial but not essential) Episomes – plasmids that become incorporated into genome Genetic Variation in Bacteria Conjugation – exchange of DNA through pilus (genome or plasmid) Transduction – virus introduces new DNA to bacterial genome (virus picked up DNA from another bacteria) Transformation – bacteria absorb DNA from surroundings and incorporate into genome Recombinant DNA DNA segments or genes from different sources Natural: transduction, conjugation, transposons, crossing over Artificial: recombinant DNA technology rDNA Technology Restriction enzymes – from bacteria (defense against viruses), cut DNA at specific sequences Uneven cuts produce “sticky ends” GOI = gene of interest Vector = method of introducing foreign DNA into cell (usually plasmid) Transformation - uptake of vector containing GOI cDNA Problem: GOI contains both exons and introns (introns can suppress expression) Solution: copy DNA from already processed mRNA using viral reverse transcriptase Called complimentary DNA (cDNA) PCR Polymerase Chain Reaction Used to amplify DNA (make a LOT of copies of a sample) Uses DNA polymerase directly (and synthetic primers) RFLP Analysis Restriction Fragment Length Polymorphisms Polymorphism = variation in DNA sequences between individuals Restriction enzymes from bacteria cut DNA molecules to create DNA fragments of varying lengths Separate fragments in gel electrophoresis DNA fingerprinting (CSI, paternity) Bozeman: DNA Fingerprinting DNA Fingerprinting http://youtu.be/DbR9xMXuK7c (6:10)