BIOL 675 8/24, 8/26, 8/28 Notes
BIOL 675 8/24, 8/26, 8/28 Notes 675
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This 5 page Class Notes was uploaded by . Notetaker on Friday August 28, 2015. The Class Notes belongs to 675 at Kansas State University taught by Revathi Govind in Fall 2015. Since its upload, it has received 69 views. For similar materials see Genetics of Microorganisms in Biology at Kansas State University.
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Date Created: 08/28/15
Genetics of Microorganisms 675 82415 I Why study microorganisms a Earth sustains life because of microbes b Central dogma DNA replication transcription RNA translation proteins i DNA nucleotides ii Proteins amino acids ll Carbon cycle a C02 in atmosphere photosynthesis add sunlight organic carbon decay organisms animal or plant respiration or fossils and fossil fuels from thee dead organisms and waste products lll Nitrogen cycle a Nitrogen into ammonia nitrobacters x into nitrites nitri cation into nitrates plant consumption or gaseous losses b Decomposition of Organic matter i Saprophytes tissues into proteins ii Fermentative bacteria proteins into amino acids iii Acetogens amino acids to acetic acid N2 iv Methanogens acetic acid to C02 methane IV Human civilization a Microorganisms have signi cant socialeconomic impact i Human diseasecausing agents 1 Plague in 14th century 3040 population Bubonic through lymph nodes Pneumonic lung Systemic blood Alexandre yersin swiss physician found it39s a bacteria in the lymph node and told Pasteur it was gram negative 2 Irish famine 174041 40 irish population wiped out 3 Global warming do cows pollute as much as cars a 100500 gallons of methanecowday ii Microbes in industry 1 Fermentation along with fungi used to prepare cheese soy sauce yogurt 2 Mining pyrite ore bacteria breaks this ore and releases dissolved metals and is separated by electrophoresis 3 Bioremediation by oileating microbes 4 Diseases V Why study the genetics a They are important model systems i Convenient to grow in large quantities ii Grow rapidly iii Mostly single celled less complex haploid iv Basic cellular processes similar to higher organizations Vl Terminology a Prokaryote before nucleus no internal membrane bound structures i Divisions eubacteria amp archaea ii Archaea single celled microbes apem 1 Extreme environments high temp high S low pH high salt 2 Methanogens 3 Some processes closer to eukaryotes b Eukaryote true nucleus internal structures c Horizontal gene transfer info from one species into another species i May cause antibiotic resistance virulence factors ii Ex e coli ancestor gave rise to shigella by two chromosomal regions and plasmid deletions in two genes Vertical info from parent to offspring e Notation i Insertion mutation trpETn10 tn10 is a transposon 1 When it is capitalized TrpE it39s a protein ii Deletion mutation deltatrpE iii Example from class 1 Eco DH5alpha strain genotype 2 endA1 recA1 deoR hst17 rK mk Genetics of Microorganisms 675 82615 0 l DNA structure a First the history began with biological function then broke off into geneticists amp biochemists i Geneticists want to know the quotwholequot organisms produce mutants understand function by mutants vs organisms ii Biochemist fractionate everything break into compounds and compare each compound iii Molecular biology is born putting the two elds together to retain the whole picture b What led to the discovery i Frederick Griffith 19205 British screen for bacterial strains that came from pneumonia streptococcus pneumonia 1 He found some strains were making small colonies but some were making large colonies 2 Injected these two strains in animals rough strain survive smooth strain dies smooth strain makes sugar capsules and immune system cant detect the strain a Heat killed smooth strain mouse lives b Mixed heat killed smooth strain amp rough strain mouse dies ii Transforming principle iii Oswald avery boss man 1 Two colleagues repeated the Grif th experiment wo animals a Fractionated the smooth form into proteins and nonproteins and incubated with rough the non protein mixture transformed the info DNA iv Chase amp Hershey what is DNA made of 1 Used e coli amp incubated with bacteriophages some info from phages was being inserted into e coli a Experiment details i You need ecoi proteins marked phage DNA marked phage ii Label DNA with phosphate iii Label proteins with Sulfur iv Put phage in bacterial cells blender to remove phage centrifuge to separate phage from e coli v Erwin chargaff rst to report ATCG are unique vi Watson amp crick two strands complimentary double helix c Structure i Nucleotides form repeating units 1 Phosphate sugars bases purines pyrimidines 2 Base sugar nuceoside 3 Base sugar phosphate nucleotide ii Phosphodiester bonds link nucleotides to form strand iii Two strands interact to form helix iv Deoxyribonucleotide precursor synthesis 1 ADP ribonucleotide reductase deoxyribose dADP kinase dATP 2 UDP dUDP dUTP phosphatases dUMP thymidylate synthase dTM P d39lTP v Nucleotide polymerization reaction phosphodiester bond formation 1 ln nucleotides 1 base 5 phosphates 2 Phosphate in 539 reacts in 339 OH molecule 3 DNA ALWAYS GROWS IN THE 339 end vi When DNA strands come together connect with H bonds 1 3 GC 2 2AT 3 Melting point is proportional to GC 4 Melting curve nd the middle of the curve and look at the x axns Genetics of Microorganisms August 28 2015 l DNA replication Replication starts at a unique site ori or oriC Bidirectional Polymerization proceeds by 539 to 339 addition of dNTP Semi conservativethe two new strands have half of their parent strand i Conservative only one of the two new strands has information from the parent ii Dispersive the two new strands and information dispersed within the two new strands from the parent DNA iii Experiment to prove this take home assignment 1 Objectives materials method procedures results conclusion apem e Semidiscontinuous means the two mother strands are used as a template to the DNA polymerase i Growth of leading strand continuous on the 539 strand ii Lagging growth towards the 339 end not continuous f lnitiation origin of replication in E coli is termed oriC i Three types 1 Atrich region easily break bonds 2 DnaA boxes a dnaA protein binds the boxes also recruits other histone like proteins to bind the result is the AT region opens up and now the junction is made which is the replication fork i on the fork dnaB and dnaC binds B helicase C bring in the helicase to the fork carrier b bidirectional replication happens on both sides of the fork c DnaB uses ATP for energy only have ATP on 4 sides of the protein on the side it doesn39t have ATP the monomer is charged and it moved to the next monomer site this is how the protein moves g Singlestranded binding protein helix destabilizing protein i Sometimes regions rich is A and regions rich in T form loops in the DNA strand the polymerase is UNABLE to go on these regions 1 Solution single stranded binding proteins bind on the strand and quotstraightenquot the strand out h Primase makes primers short RNA oligonucleotides that DNA polymerase extends i dnaG primase or chromosomal replication i DNA polymerase adds bases from 539 to 339 j New mode trombone model i Polymerases are connected with one another ii Imagine the template is food the leading strand has food in front of him the lagging strands food is behind him he needs proteins to put the DNA in front him to eat 1 Beta clamp binds the polymerase and activates the lagging polymerase to work 2 DNA polymerase III is responsible for the replication a Components i epsilon component dnaQ proofreads the sequence moves 339 to 539 when they are mistakes ii tau protein binds polymerases together iii topoisomerases relax tension in supercoiled DNA that results from DNA helicase action resolve daughter molecules at the end of replication 1 topA cuts DNA strand moves DNA down and now its one twist less connect it back 2 type II gyrA gryB relaxes supercoils cuts both strands of dNA and it goes around one coil and connects them back cut again repeats k DNA polymerase I removes RNA primers and then DNA ligase comes and joins 5quot P04 and 3quot OH to seal nicks l Identi cation of genes for DNA replication i E coli culture created mutants