IntroductiontoGenetics

Sunday,February14,2010 11:17AM

A. GeneticsIsImportant
a. Genesinfluenceslivesinsignificantway andarefundamentaltowhoandwhatweare. i. Inagriculture:Thehereditaryofnaturehasbeenunderstoodforthousandyears(genetic alteration/domesticationofplantsandanimals) ii. Inpharmaceuticalindustry:synthesisofdrugs,growthhormone,insulinfromfungiand bacteria.Biotechcompaniesproducerecombinantproteinsinbioreactors iii. Moleculargeneticshavebeenusedtoproducebacteriathatremovemineralsfromore,break downtoxicchemicalsandinhibitdamagingfrostformation iv. Inmedicine:discoveryofhereditarydiseasesanddisorders(somecancers,sicklecellanemia etc.) v. Inbiology:unifyingprinciple similargeneticsystemamongorganisms b. Divisionofgenetics: transmissiongenetics (howtraitsarepassed),moleculargenetics (chemical natureofthegene)andpopulationgenetics (geneticcompositionofgroups) c. Modelgeneticorganism Drosophila melanogaster (fruitfly) Escherichia coli (bacterium)

Caenorhabditis (roundworm)
Mus musculus (housemouse)

Arabidopsis thaliana (thalecressplant) Saccharomyces cerevisiae (baker'syeast)

Shortgenerationtime,manageablenumberofprogeny,adaptabilitytolabenvironment,the abilitytobehousedandpropagatedinexpensively

B. Humans'UnderstandingofGenetics
a. Pangenesis: specificparticles(gemmules)carryinformationfromvariouspartsofthebodytothe reproductiveorgans,fromwhichthearepassedtotheembryoatthemomentofconception.Lead totheinheritanceofacquiredcharacteristics b. Preformationism: insidetheeggorspermthereexistsatinyminiatureadult(homunculus), inheritingalltraitsfromeitherthefatherorthemother c. Blendinginheritance: offspringareamixtureofparentaltraits d. Germplasmtheory: thecellsinthereproductiveorganscarryacompletesetofgeneticsinformation thatispassedtotheeggandsperm e. GregorMendel: basicprinciplesofinheritance f. JamesWatsonandFrancisCrick: 3DstructureofDNA g. TheHumanGenomeProject: launch1990.In1995,thefirstcompleteDNAsequenceof Haemophilusinfluenzaewasdetermined.1996,yeast. h. Thefutureofgenetics: i. Geneticmicrochips:activityofthousandsofgenesinagivencell ii. Proteomics:modelofthestructuresandfunctionsofproteinscomputerizedfromDNA sequence iii. Personalizedgeneticmedicine:assesstherisksofacquiringvariousdiseasesandtailortheir treatment

C. FundamentalConcepts
a. b. c. d. Cellsareoftwobasictypes:eukaryoticandprokaryotic Thegeneisthefundamentalunitofheredity Genescomeinmultipleformscalledalleles Genesconferphenotypes.Traitsarenotinheriteddirectly,genesare.

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e. f. g. h. i. j. k.

GeneticinformationiscarriedinDNAandRNA Geneticarelocatedonchromosomes. Chromosomesseparatethroughtheprocessesofmitosisandmeiosis GeneticinformationistransferredfromDNAtoRNAtoprotein Mutationsarepermanent,heritablechangesingeneticinformation Sometraitsareaffectedbymultiplefactors Evolutionisgeneticchange

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CellularReproduction
Saturday,February13,2010 12:06PM

Prokaryoticcells Unicellular Simplecellstructure Lackanucleus CircularDNAwithnohistone.

Eukaryoticcells Unicellularormulticellular Compartmentalizedcellstructurewithmembrane boundcomponents Membraneboundnucleus Histonesallowtightpackagingoflongstrandsof DNA(Hist+DNA=chromatin>chromosome)and limittheaccessibilityofenzymes

A. CELLREPRODUCTION
ProkaryoticCellReproduction Startsattheoriginofreplication Insomebacteria,proteinsbindthereplicationoriginsandanchorthenewchromosomes totheplasmamembraneatoppositeends EukaryoticCellReproduction 1. EukaryoticChromosomes Eachspecieshasacharacteristicnumberofchromosomespercell.Nosignificance betweenthecomplexityofanorganismanditsnumberofchromosome Inmosteuk.cells,thereare2setsofchromosomesasaconsequenceofsexual reproduction. Twochromosomesinahomologouspairareusuallyalikeinstructureandsize(copiesof thesamegeneon2chr.ofahomo.Pair:alleles) Mostcellshave2setsofgeneticinformation:diploidcells Spermcellsandeggcellsonlyhave1set:haploidcells 2. ChromosomeStructure Largeandcomplexrodshape DNAsarehighlyfoldedandcondensed,coiledaroundhistones Thinanddifficulttoobservemostofthetime Condensebeforecelldivision(Prophase) 3elements: Centromeres:attachptforspindleMTsthroughkinetochoreproteincomplex.Lack ofcentromere>chromosomesarelost.Fortype:metacentric,submetacentric, acrocentricand telocentric Telomeres: tipsofchromosomes,stabilizetheends. Originsofreplication: siteswhereDNAsynthesisbegins.Twoinitiallyidentical copiesofchromosome:sisterchromatids

B. CELLCYLEANDMITOSIS
Thecellcycleisanorderedseriesofprocessesthatthecellpassesthroughonitswaytodivision Therearevariouscheckpointspresentinthecycletoinsurethefidelityofthecellcycle G1/Scheckpoint: holdsthecellinG1 untilithasallthenecessaryenzymeforDNA replication.Beforereachingthispoint,thecellmayenterG0phase constantsize G2/Mcheckpoint: checkforundamagedDNA Spindleassemblycheckpoint (duringmetaphase):checkifallchromosomesareattached tospindlefibersandalignedonthemetaphaseplate.

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1. Cellcycle Interphase growthandpreparationformitosisanddivision G1phase:cellsgrow,proteinsnecessaryforcelldivisionaresynthesized.Cellsmay enterandremaininG0 Sphase:chromosomesduplicate.AfterSphase,eachchr.iscomposedof2sister chromatids G2phase:addtlbiochemicalevents Prophase replicatedchromosomescondense,mitoticspindleforms Prometaphase nuclearenvelopedisintegrates,spindlemicrotubulesform Metaphase chromosomesarealignedonthemetaphaseplate Anaphase sisterchromatidsseparateformingindividualchromosomes Telophase chromosomesarriveatspindlepoles,nuclearenvelopereforms, chromosomesdecondense 2. Consequencesofcellcycle: Twocellsthatcontainthesamegeneticinfoasthemothercell Nonetreductionorincreaseinchromosomenumber Eachcellcontainshalfthecytoplasmandorganellescontentoftheparentalcell Resultingcellsaregeneticallyidenticalbutnotidenticalincytoplasmiccontent

C. SEXUALREPRODUCTION
1. Meiosis: consistsoftwodivisions MeiosisI ProphaseI: Chr.Condense,homologouschromosomesynapseintobivalent/tetrad, crossingovertakesplace,nuclearenvelopbreaksdown,mitoticspindleforms MetaphaseI: homologouspairsofchromosomeslineuponthemetaphaseplate AnaphaseIP homologouschromosomes(eachwithtwochromatids)separateand movetowardoppositepoles TelophaseI chromosomesarriveatthespindlepoles Cytokinesis: thecytoplasmdivides,producing2cells,eachhavinghalfoftheoriginal #ofchromosomes Interkinesis: insometypesofcells,thespindlebreaksdown,chromosomesrelaxanda nuclearenvelopreformsbutnoDNAsynthesistakesplace MeiosisII ProphaseII (onlyforcellsthathaveinterkinesis):chromosomescondense,the spindleforms,nuclearenvelopdisintegrates MetaphaseII:individualchromosomeslineuponthemetaphaseplate AnaphaseII:sisterchromatidsseparateandmoveasindividualchromosome towardthespindlepoles TelophaseII:Chromosomesarriveatthespindlepoles,thespindlebreaksdownand anuclearenvelopreforms Cytokinesis: thecytoplasmdivides 2. Consequencesofmeiosis Eachdiploidoriginalcellproduces4haploidcells Chromosomenumberisreducedbyhalf Cellsproducedbymeiosisaregeneticallydifferentfromoneanotherandfromthe parentalcell geneticvariation CrossingoverinprophaseIshufflesallelesonthesamechromosome referstotheexchangeofgenesbetweennonsisterchromatids(fromdifferent homologouschr).

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Aftercrossingover,sisterchromatidsarenolongeridentical.Newcombinationsof allelesonachromatids(intrachromosomalrecombination) RandomdistributionofchromosomeinanaphaseIshufflesallelesondifferent chromosomes

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BasicPrincipleofHeredity
Saturday,January30,2010 7:32PM

1. MonohybridCrossesRevealsthePrincipleofSegregationandtheConceptofDominance a. Principleofsegregation i. Eachindividualdiploidorganismpossessestwoallelesforanycharacteristic ii. Thesetwoallelessegregatewhengametesareformed,oneallelegoesintoeachgametes iii. Twoallelessegregatewithequalprobability iv. Dominance:whentwodifferentallelearepresentinagenotype,onlythetraitencodedby oneofthem the"dominant"allele isobservedinthephenotype 2. Testcross:individualofunknowngenotypeiscrossedwithhomozygousrecessivegenotype 3. IncompleteDominance a. Theheterozygotehasaphenotypeintermediatebetweenthephenotypesofthetwohomozygote 4. DihybridCrossesRevealthePrincipleofIndependentAssortment a. PrincipleofIndependentAssortment i. Allelesatdifferentlociseparateindependentlyofoneanother ii. Applytocharactersencodedindifferentchromosomes

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SexDetermination
Friday,February12,2010 1:16PM

Armadillidium vulgare (isopods):geneticmalesmaybeconvertedintofunctionalfemalesbythe presentsofinfectingbacteria Inmammalsandhuman: sexisdeterminedbythepresenceofsexchromosomes:females possess2XchromosomesandmalespossessoneXandoneY Insomeotherorganisms (e.g.A.vulgare)thesituationisreversed:femaleZW,maleZZ Sexualreproduction: theformationofoffspringsthataregeneticallydifferentfromtheirparents 2processes:meiosisproduceshaploidgametesandfertilizationproducesdiploidzygotes Sexualphenotypes:maleandfemale.Malesproducesmallgametes;femalesproduce largegametes Hermaphroditism:bothsexesarepresentinthesameorganism(monoecious).Organismshas eithermaleorfemalereproductivestructures(dioecious)

A. MechanismsofSexDetermination
a. Sexchromosomes: i. Xchromosome:larger;metacentric ii. Ychromosome:smaller;acrocentric. 1) 2/3isheterochromatin:consistingofshortrepetitiveDNAsequences(no activegenes) 2) 1/3iseuchromatin:fewgenes iii. XandYarehomologousonlyatpseudoautosomalregions,allowingthemtopairup inmeiosis iv. Platypus:femaleshave5pairsofXandmaleshave5pairsofXandY b. Chromosomalsexdetermination i. Obviousdifferencesbetweensexchromosomes ii. Theindividualgeneslocatedonsexchromosomesdeterminethesex,notthe presenceofthesexchromosomethemselves.(SRYgeneonYchr.determinesmale phenotype) Type XXXO Female XX Homogametic:All gameteshaveX Male XO Heterogametic:1/2 gameteshaveX;1/2 gameteshavenone XY Heterogametic Examples Grasshopper; C.elegans

XXXY

XX Homogametic

Human;all mammals; someinsects, reptiles,plant Birds,snakes, butterfliesand some amphibians andfishes; isopods

ZZZW

ZW Heterogametic

ZZ Homogametic

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Haploidiploidy Nosexchr. Sexisbasedon numberofchr. sets

Developfromfertilized eggs Twosetsof chromosomes Produceeggsbymeiosis Sisters:50%frommom, 100%fromdad,75% related.

Developfromunfertilized Insectsinthe order eggs Singlesetofchromosomes Humenoptera Producespermsbymitosis Brothers:50%frommom, 50%related

c. Genicsexdetermination (insomeplantsandprotozoans) i. Noobviousdifferencesbetweensexchromosomes(nosexchr.),sexisdetermined genetically:genotypesatoneormorelocideterminethesex d. Environmentalsexdetermination i. MarinemolluskCrepidulafornicata(slipperlimpet) sequentialhermaphroditism. 1) Firstlavatosettledevelopintoafemalelimpet,producingchemicalsto attractotherlarvae,whichthensettleontopofit.Theselarvaedevelopinto malesandserveasmatesforthelimpetbelow. 2) Afteraperiodoftime,themalesontopdevelopintofemalesandattract additionallarvae ii. Turtle:warmtemperaturesduringembryonicdevelopmentproducefemale whereascooltemperaturesproducemales iii. Crocodilians:reverse e. SexdeterminationinDrosphilamelanogaster i. 8chromosomes:3pairsofautosomesandonepairofsexchr. ii. Normally,XXXY:femaleXX,maleXY iii. ThepresenceofYchr.doesnotdeterminemaleness. iv. Genicbalancesystem:sexisdeterminedbyabalancebetweengenesonautosomes andXchr. 1) Autosomes:geneswithmaleproducingeffects 2) Xchromosomes:geneswithfemaleproducingeffects 3) X:Aratio thenumberofXchromosomesdividedbynumberofhaploidsets ofautosomes X:A>1 X:A=1 X:A=0.5 X:A<0.5 Metafemale Female Male Metamale

0.5<X:A<1 Intersex

B. Sexdeterminationinhumans
a. Theroleofsexchromosomes i. Xchr.containsgeneticinformationforbothsexes(onecopyisrequired) ii. SRYgenelocatedonYchrdeterminesmaleness.AbsenceofY>female iii. GenesaffectingfertilityareontheXandY.FemalesneedtwoX'stobefertile iv. AddtlcopiesofXmayupsetnormaldevelopmentinbothmalesandfemales b. Maledetermining i. SexdeterminingregionY(SRY)geneencodesaproteinthatbindstoDNAand causesasharpbendinthemolecule,causingtheneutralgonadstodevelopsinto

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testes ii. TestessecrettestosteroneandMullerianinhibitingsubstance 1) Testosteroneinducesthedevelopmentofmalecharacteristics 2) Mullerianinhibitingsubstancecausesthedegenerationoffemale reproductiveducts iii. SRYalsoplayaroleinfertilityandthedevelopmentofsexdifferences c. Androgeninsensitivitysyndrome i. FemalecharacteristicandpsychologicalorientationbutXY ii. Vaginaendsblindly,uterus,oviductsandovariesareabsent iii. Testeslieinsidetheabdominalcavity,producingmaleleveltestosterone iv. Defectiveandrogenreceptorsmakecellsinsensitivetotestosterone v. GeneforandrogenreceptorisonX,inheritedfromthemother d. RSPO1mutation: RSPO1promotesthedevelopmentofgonadstoovary e. Turnersyndrome(XO) female(1/3000) i. Normalintelligence,underdevelopedsecondarysexcharacteristics ii. Shortandhavelowhair,relativelybroadchest,foldsofskinontheneck iii. Highrisksofadultdiseases,sterile f. Klinefeltersyndrome (XXY XXXY XXXXY XXYY) male(1/1000) i. Normalintelligence,tallerthannormal ii. Smalltestes,reducedfacialandpublichair iii. Sterile g. TriploXsyndromes (polyX) female i. Greaterchanceofretardation,mostXXXhavenormalintelligence ii. Tallandthin

C. SexlinkedCharacteristics
Determinedbysexchromosomes MostareXlinkedandfewareYlinked Xlinked a. ForXlinkedtrait,maleishemizygousbecauseonlyonealleleispresent b. Morgan'sexperiment onXlinkedwhiteeyetraitinDrosophila i. CalvinBridge:furtherexperiment ii. Nondisjuction:10%ofthetime,twoXchromosomesfailtoseparateinanaphaseI c. XlinkedRGcolorblindnessinhumans i. Conecellsthatlinetheretinacontainoneofthreepigmentscodedondifferentloci: blueonchr.7,redandgreenon X ii. Affectedmompassesthetraittohersonsbutnotherdaughter iii. Affecteddadpassesthetraittohisgrandsonthroughhisdaughterbutnevertohis son. d. Dosagecompensation i. TheamountofproteinsencodedbygenesonXwouldbetwiceinfemalesthanin males ii. Dosagecompensationequalizesthisamountinmalesandfemales 1) Fruitflies:doubletheactivityofXinmale 2) C.elegan:halvetheactivityofXinfemale 3) Placentalmammals:inactiveoneoftwoXchromosomes iii. Dosagecompensationinhumans/mammals

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1) BytheactionofXinactivationspecifictranscript(Xist) geneonXchr.Xist encodesanRNA(notaprotein)thatcoatstheXchr.andinducesinactivation. XististheonlygeneoninactiveXthatisexpressed. 2) Barrbody: randomlyinactivatedXchromosome(s) 3) OnlyoneXchromosomeisactive 4) Femalesarefunctionallyhemizygous:50%cellsexpressonealleleand50% expresstheotherallele.Cellsinfemalearenotidenticalwithrespecttothe expressionofthegenesontheXchromosomes. Femalesare"mosaics" 5) RandomXinactivation takesplaceearlyindevelopment.Afterbeing inactivated,anXchromosomeremainsinactiveinallsomaticcellsthat descendfromit 6) Example:tortoiseshellcats Malescanonlybeeitherblackororange;females maybeblack,orangeorpatchymixtureofblackandorange 7) IfonlyoneXchromosomeisactiveinonecell,thepresenceofmultipleX's shouldnotbedetrimental.Infact(casseofXO,XXY), someXlinkedgenes (15%)completelyescapeinactivation,other10%inactiveinsome(butnot all)females Zlinked a. ThesameasthatofXlinkedcharacteristics,exceptthatthepatterninmalesandfemales isreverse Ylinked 8palindromesequencesallowYchromosometorecombinewithitself

a.

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PedigreeAnalysis
Thursday,February11,2010 9:37PM

1. ConstraintsontheStudyofGeneticsinHuman Controlledmatingarenotpossible Longgenerationtime Humanfamilysizeissmall

2. PedigreestoStudytheInheritanceofCharacteristicsinHumans a. Pedigree: schematicdiagramofafamilyhistory,oftenwithphenotypicandgenotypicdata. b. Waardenburgsydrome: autosomaldominant,deafness,fairskin,whiteforelock,visual problems c. Proband: thepersonfromwhomthepedigreeisinitiated

3. PedigreeAnalysis a. Autosomalrecessive: Appearonlyifapersoninheritstwoallelesforthetrait Equalfrequencyb/tmaleandfemale Tendstoskipgeneration Morelikelyincaseofconsanguinity(greaterchanceofbothpossessingtherarerecessive allele) Affectedoffspringareusuallyborntounaffectedparents E.g:TaySachsdisease:healthyatbirthbutbecomelistlessandweak.Physicaland neurologicalconditionsworsen,leadingtoblindness,deafness,deathat23age b. Autosomaldominant Atleastonealleleisinheritedfromoneparent Equalfrequency Donotskipgenerations Unaffectedpeopledonottransmitthetraittotheirdescendants Affectedoffspringmusthaveanaffectedparent(unlessmutation) E.g:hypercholesterolemia adefectonchromosome19thatleadstoaverylowlevelof thelowdensitylipoprotein(LDL)receptor.Bloodcholesterolisgreatlyelevated c. Xlinkedrecessivetraits Morefrequentlyinmalesthaninfemales,becausemaleneedinheritonlyasinglecopyof thealleletodisplaythetrait Affectedsonareborntounaffectedmotherswhocarrythegene.(Onesonisaffected thenmomcarriesthegene) Tendstoskipgeneration(unaffectedfemaletoaffectedmale Notpassedfromfathertoson,becauseXisalwaysfromthemother E.g:hemophiliaA,absenceofaproteinnecessaryforbloodtoclotdueto abnormal/missingfactorVIIIlocatedonthetipofthelongarmoftheX d. Xlinkeddominanttraits Affectmorefemalethanmale PersonwithXlinkeddominanttraitsmusthaveanaffectedparents Donotskipgenerations

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Notpassedfromfathertosonbutfromfathertoalldaughters.Affectedsonsmusthave affectedmother E.g:hypophosphatemia(familialvitaminDresistantrickets),resemblerickets:bone deformities,stiffspinesandjoints,bowedlegs,andmildgrowthdeficiencies.Large phosphateamountinurineandlowinblood,reducingdepositionofmineralsinthebone e. Ylinkedtraits Onlymalesareaffected Affectedfatherspassthetraittoallsons Donotskipgenerations 4. ImportanceofGenesvsEnvironmentonVariationinaTrait a. Twinstudy Monozygotictwins:1egg+1sperm Dizygotictwins:2eggs+2sperms Share100%genes Share50%genes Frequencyvariesamongpopulation Frequencydoesnotvary(4/1000) andwithmaternalage(higherwith higherage) Tendstoruninfamilies Littletendencytoruninfamilies Concordant:bothmembersofatwinpairhaveatrait Discordant:onehasthetrait,onedoesn't Concordance:thepercentageoftwinpairsthatareconcordantforatrait Geneticallyinfluencetraitsshouldexhibithigherconcordanceinmonozygotictwins Highconcordanceinmonozygotictwinbyitself(i.e.thatindizygotictwinsisashigh)does notsignalageneticinfluence ThomasBouchardandNancySegalpioneeredstudiesdealingwithMZandDZtwins raisedapart b. Adoptionstudy Adoptedchildrenshare50%geneswiththeirbiologicalparentsandsharethesame evironmentwiththeiradoptiveparents Ifadoptedpersonsshowsimilaritiesinacharacteristicwiththeiradoptiveparents,these similaritiesareduetoenvironment Ifadoptedpersonsshowsimilaritiesinacharacteristicwiththeirbiologicalparents,these similaritiesareduetogenes 5. GeneticTesting:Recognizedthepotentialforageneticconditionatanearlystage a. Untrasonography Highfrequencysoundisbeamedintotheuterus,whenthesoundwavesencounter densetissue,theybouncebackandaretransformedintoapicture b. Amniocentesis Obtainingasampleofamnioticfluid,whichcontainsfetalcells Ultrasonographyisusedtomonitor,alongneedleisinsertedthroughabdominalwall intotheamnioticsactowithdrawnamnioticfluid Fetalcellsareseparatedandcultured,whichtakestime Usedtoanalyzechromosomecomplement c. Chorionicvillussampling(CVS)

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Obtainingasmallpieceofthechorioncomposedoffetalcells Acatheterisinsertedintothevaginaandispushedthroughthecervixintotheuterus, thetipisplacedintocontactwiththechorion,suctionisthenapplied Fasterthanamniocentesisbecausecellsdon'thavetobecultured d. Maternalbloodtests Candetectsomegeneticconditionssuchasonesrelatedalphafetoproteinlevelinthe mother'sblood e. Noninvasivefetaldiagnosis FetalcellsandfreefloatingfetalDNAinmaternalbloodareseparatedandtested DetectDownsyndromeandcysticfibrosis NotasaccurateasamniocentesisorCVSandonlymutatedgenesfromthefathercanbe detected f. Preimplantationgeneticdiagnosis(PGD) Canbeusedinconjunctionwithinvitrofertilizationtechniquestoanalyzeanembryofor geneticabnormalitiesbeforeimplantingtheembryo Onecellisremovedfromthe8 or16cellembryostoanalyze Healthyembryoisselectedandimplanted 6. PostnatalGeneticTesting a. Newbornscreening: phenylketonuriaandgalactosemiaifnottreatedatearlyagecanresultin mentalretardation b. Heterozygotescreening:toidentifyheterozygouscarriers(e.g.TaySachs) c. Presymptomatictesting: evaluatinghealthypeople

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