Week1/2: ClassicalGenetics GenesInheritance,andUs.

LearningOutcomes(Lecture2)
Studentsshouldunderstandthelandmark findings(andpeople)thatcontributedto modernbiologyandbiotechnology. Studentsshouldcomprehendthatcellsare thebasic,autonomousunitsoflife andthat DNAcontainstheinformationtosustainlife. Studentswillunderstandhowthe information responsibleforcelldiversityis storedinthechemicalformofDNA.

LearningOutcomes(Lecture3)
Studentsshouldbeabletounderstandwhatare heritabletraitsandhowtheyareinherited(e.g. meiosis,mitosis). UsingMendelian genetics(e.g.Punnetsquare), studentsshouldbeabletoappreciateandpredict howinformationismonitoredovertime. Studentswillhaveanideaofhowalterationof informationarises (e.g.chromosomalaberrations) andhowitcancontributetoheritabletraits.
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OUTLINE:KeyConcepts/Points
ClassicalGenetics:inheritanceandtraits (slides6 24) Whatandwhereisthegeneticmaterial? (slides25 46) Hereditarytraits&celldivision: mitosis&meiosis (slides48 60) Chromosomedynamics: normalvs aberrations (slides61 73)

1832caricaturetitled"AChipoffthe OldBlock".

Whatcanorcantbe inherited?Somehuman traitswereincorrectly thoughttobeinherited.

http://www.dnalc.org/

Genotype & Phenotype Description http://www.dnalc.org/resources/dnatoday/110210-genotype-phenotype.html

1865 Gregor Mendel

FirsttoproposePaired HereditaryFactors andBasicLawsofHeredity

AbbeyofSt.ThomaswhereMendeldidhis earlygeneticpeastudies

Took8yearsandmorethan10,000peaplants!

www.dnalc.org
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Seven pairs of contrasting traits and the results of Mendel's crosses of the garden pea (Pisum sativum). In each case, pollen taken from true-bred plants exhibiting one trait was used to fertilize the ova of true-bred plants exhibiting the other trait. In the F1 generation, only one of the two traits was exhibited by all plants. The contrasting trait reappeared in approximately 1/4 of the F2 plants (progenies of F1).

Genetic characters are controlled by discrete factors that exist in pairs in individual organisms. A discrete paired-factor exists for each trait.

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Thediscretefactorthat controlsthetextureofthe peaisthetexturegene.

Sexcells
(containonlyoneof thediscretefactor fromthepair)

Alternateformsofthe texturegene,whetherS or

s,arereferredtoas alleles.
Allelesarealternate formsofagene

Dominantvs recessivetraits
SeeIVLEanimation&CD ROM
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1:2:1

Gregor MendelsBasicLawsofHeredity

Gy Gory

Greenyellowallelehybrids
(F1parents) (Sexcells)

Gy Gory

F2offspring:

GG

Gy

yG

yy

Ratioof1:2:1forGG:Gy :yy
NOTE:AlloffspringhaveTWOallelesforthesamecharacteristic (colour),onefromeachparent. 13

GG:Gy :yy
Thesearethethreegenotypes
Genotype:Thegeneticconstitutionorgeneticmakeupofacelloran individualorganism;itcanpertaintoone,ormorethanonegene(s).A homozygousgenotypehasthesamealleles;aheterozygousgenotypehas differentalleles.

GG
homozygous

Gy
heterozygous

yG

yy
homozygous

Thereareonlytwophenotypes:greenoryellow
Phenotype:Theobservablephysicalorbiochemicalcharacteristicsofan organism,asdeterminedbyBOTHgeneticmakeupANDenvironmental influences.
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MendelsThreeLawsofHeredity:
1. LawofSegregation:
Genetictraitsarecontrolledbypairedunitfactors(later knownasgenes);oneform(allele)ofeachpairedunitfactoris segregatedintothesexcell(e.g.alleleforyellowORgreen colour oftheseed).

2.LawofDominance:
Oneformofthegene(allele)willalwaysbedominant.

3.LawofIndependentAssortment:
Differenttraitsareinheritedindependentlyfromeachother (e.g.canbetallandgreenortallandyellow). Note:Mendelslawsarenotalwaystrueinnature.
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Dominance&Recessiveness
Incomplete/PartialDominance & Codominance

16

DominantTraitandAllele
Dominanttraitreferstoageneticfeaturethathides therecessivetraitinthephenotype ofanindividual.A dominantallelecausesitsphenotypetobeseenina heterozygous(Aa)genotype. Forexample,ifapersonhasonealleleforblueeyes andoneforbrown,thatpersonwillalwayshave browneyesbecausebrownisthedominanttrait.For apersontohaveblueeyes,boththeirallelesmustbe blue(recessive).Whenapersonhastwodominant alleles,theyarereferredtoashomozygousdominant. Iftheyhaveonedominantalleleandonerecessive allele,theyarereferredtoasheterozygous.
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Howisthispossible?
Dr X
sperm

Dr

egg

D
DD Dr

r
rD rr

D r

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RecessiveTraitandAllele
Theterm"recessiveallele"referstoanallelethatcausesa phenotype(visibleordetectablecharacteristic)thatisonly seeninahomozygousgenotype (anorganismthathastwo copiesofthesamerecessivetrait)andneverina heterozygousgenotype. Everypersonhastwocopiesofeverygeneforeachpairof autosomal(nonsexdetermining)chromosomes;onegeneis onthechromosomefromtheirmotherandoneisonthe chromosomefromtheirfather.Forrecessivegenetictraits,a personMUSTinherittwocopiesoftherecessivegeneforthe traittobeobserved.Thus,bothparentshavetobecarriers ofarecessivetraitinorderfortheiroffspringtoexpressthat trait.Whenparentsarecarriers,thereisa25%chancethe recessivetraitwillbeobservedwitheachoffspring.
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Modificationto Mendelian Ratios R1R1 x

red
R1R2 pink

R2R2 P1 white
X
homozygousparent homozygousparent

F1

F1 offspring heterozygous forflowercolor

Incomplete dominance:
Onealleleofapairisnot fullydominantoverits partner,sothephenotype oftheheterozygoteis somewhereinbetweenthe phenotypesofthetwo parents. F2 offspring 3phenotypesin 1:2:1ratio SeeIVLEanimation

R1R2

xR1R2

F1 xF1

R1R1 red R1R2 pink F2 R2R2 white

MultipleAllelesABOBloodGroupInheritance(Aand BareCodominant;Oisrecessive)
Genotype AA AO BB BO AB OO Antigen (Sugar) A A B B A, B Neither (no sugar) Phenotype Full effect of A Full effect of A Full effect of B Full effect of B Full effect of AB O (no effect)

Importantforblood transfusionand organtransplantation

SeeIVLEanimationandLearn Genetics Heredity&Traits Genes&BloodType

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DistributionofABOphenotypesby raceorethnicity

* * * *

*
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Mechanismsofdominance
Thestatesofcompletedominanceandincomplete dominanceresultnotfromgenetics,butfrom biochemistry.
Thecompletenessofdominancedependsonlyonhow effectivetheproteinisatdoingitsjob.

Incompletedominancesimplymeansthathalfas manymolecules(intheheterozygote)arenotas effectiveasthenumberproducedbythe homozygote.Thisresultsinthedisplayofan intermediate,blendedphenotypeofthealleleswith varyingdegreesofcolorshades(orseverity)inthe heterozygotes. Codominance meansthatbothproteinsareequally effectiveinthecell(distinctphenotypesofboth allelesareobserved).
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CompleteDominance,IncompleteDominance &Codominance
Relationship Heterozygote Protein Genotype Expressed in Heterozygote Protein A and Protein a Heterozygote Phenotype Full effect of A

Complete Dominance (A is the dominant allele) Incomplete Dominance (no dominant allele) Codominance (both A and a are dominant alleles)

Aa

Aa Aa

Protein A and Protein a

Intermediate or blended half effect of A and half effect of a Full effect of A AND full effect of a
24

Protein A and Protein a

SeeIVLEanimations

Plant cell (eukaryote)

Animal cell (eukaryote)

Bacterium (prokaryote)

Whatand whereisthe genetic material?

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Checkout:Learn Genetics AmazingCell

MustSeeLearnGenetics: Tourofthebasics

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Whatarecellsmadeof?

Nucleic Acids

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Whatandwhereisthegeneticmaterial?
EvidencelinkingthenucleusandDNA asthesourceof hereditary/geneticmaterial 4setsofexperiments

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Cap Stalk
nucleus

Base
A. crenulata A. mediterranea

1.Hammerlings Acetabularia experiment Nucleus containsgenetic informationthatdirectscellular development.

http://cell.sio2.be/moyens/1.php
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2.GriffithsTransformationExperiment(1928)

HeatkilledtheScellsbutdidnotdestroytheirgeneticmaterial(includingthepartthat causesinfection).ThismaterialwastransferredfromdeadScellstolivingRcells. 30

Cellextract=DNA,RNA,Protein(otherstuff)

3.Avery&colleaguesexperimentthat demonstratedDNAisthetransformingmaterial

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3.Avery&colleaguesexperimentthat demonstratedDNAisthetransformingmaterial

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IsProteinorDNA TheGeneticMaterial?

4.Hershey &Chase experiment

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IsProteinorDNA TheGeneticMaterial?

TheHershey&Chaseexperimentaldesign usingbacteriophageandradioactivity(note:phage arenotdrawntoscale).

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Summaryofthe Hershey&Chase experiment demonstratingthat DNA,notprotein,is responsiblefor directingthe reproductionof bacteriophageT2 duringtheinfectionof E.coli.

Phosphorusarefoundonlyin viralDNA

Sulfurarefoundonlyin viralProtein

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SummaryofHershey&Chase

LabelinpelletANDnewphage =yes Labelinmedium=no

Labelinpelletand/ornewphage=no Labelinmedium=yes
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Gettingtoknowsometerms:
Achromosome isapackageofsomegeneticmaterial. 1chromosome=1verylongDNAmolecule AeukaryoticchromosomeconsistsofalonglinearDNAmoleculewrapped andcoiledaroundcertainpackagingproteins(e.g.histones). Aprokaryoticchromosomeisusually circular nakedDNAwithout wrappingandcoilingtohistones(althoughitmaybeassociatedwithother proteins). Manygenesarelocatedona chromosome. Aprokaryoticcellusuallyhasonechromosome. Aeukaryoticcellusuallyhasmorethanonechromosomeandtheyusuallyoccur inpairs(e.g.humanhas46chromosomesor23pairsofchromosomes). Agene isafundamentalphysicalunitofhereditythatcontainsallofthe informationrequiredtoproduceasinglepolypeptideproduct. 1gene=ashortsequenceofDNA(withspecificcharacteristics)foundonthe chromosome. Agenome istheentiregeneticmakeuporallofthegeneticinformation carriedbyacellorrequiredtomaketheorganism.Agenomeconsistsofall theDNA(allgenes)containedinallthechromosomes. 37

Earth

Cell

Allthecountries=thewhole genome=allgenetic informationthatmakethe organism;includesallthe chromosomes.

Country Districts Community

(e.g.Clementi, Pasir Ris)

Chromosome Chromosomefragment
(containsmanygenes)

Gene(complete
geneticinformationfor aprotein***)

People

Nucleotidebasepairs
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EukaryoticCell,Nucleus,Chromosome,DNA

TheDNA(human)whichis ~2 meters in length mustbe packagedintoanucleusthatisabout5micrometersindiameter.

39

PackingDNA:Howtofita whaleintoabathtub?
The~2meterlengthDNAmust bepackagedintoanucleusthatisabout~5micrometers indiameter.
Thisisacompressionofmorethan100,000 times!Thisis equivalentto~40miles(64km)ofthreadstuffedintoatennis ball. CompressionismadepossiblebywrappingtheDNAaround proteinspoolsmadeofhistoneproteinstoformstructures callednucleosomes,whicharethenpackedintohelical filaments. Chromatin (e.g.theDNA+proteincomplex)containsboth histoneandnonhistoneproteins.

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DNA is mostly double-stranded. Negatively-charged DNA is wound around positivelycharged histone proteins.

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SeeIVLE Multimedia Week2chromosome_coil.fv

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(Y) (X)

SexChromosomes: XX=female XY=male

onepairof duplicated chromosomes

Goto:Learngenetics Heredity&Traits Makeakaryotype
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Akaryotypeisthecompletesetof chromosomesofacellorindividualwhich isusuallyvisualizedduringacertainphase ofcelldivision.

Apairofhomologouschromosomes(onefrom amaleparentanditspartnerfromafemaleparent), eachintheunduplicatedstate.Homologous chromosomespairupduringmeiosisandareidentical withrespecttotheirgeneticlociand centromereplacement. Agenelocus (plural,loci),thelocationfora specificgeneonaspecifictypeofchromosome. Apairofalleles (eachbeingacertainmolecular formofagene)atcorrespondinglocionapairof homologouschromosomes. Threepairsofgenes (atthreelocionthispairof homologouschromosomes);samethingas threepairsofalleles. SeeIVLEanimation
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floweringplants

mammals
reptiles birds amphibians fish Echinoderms insects worms algae&fungi viruses& bacteria

Sizeofhaploidgenome(basepairs)
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HereditaryTraits&CellDivisions: mitosisvs meiosis

videoonIVLEMultimedia01.11_developing_egg_cells.fv
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Mother (Diploidparents)Father

MEIOSIS

Howarethe genespassedon?
Meiosis haploid=1 setofchromosomes

haploidegg

haploidsperm
fertilization

(2N 1N)
Maternal chromosome

diploid zygote

Paternal chromosome

Mitosis diploid=2 setsofchromosomes (2N 2N)

SeeIVLEanimation
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MITOSIS
diploid organism

MendelsLaws
X=motherchromosomes X=fatherchromosomes

Lawofsegregation ofalleles

Lawofindependent assortment

Aa

Aa

Pleasenotethat Onlyapairof Homologous Chromosome Isshownhere

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Aa DNAreplication

Aa

Pleasenotethat Onlyapairof Homologous Chromosome Isshownhere

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Aa

Aa

a a

A A

Please note that only ONE pair of homologous chromosomes is shown here

Aa

aA

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53

1911
ThomasH. Morgan
NobelPrizewinner(1933)

Formulated ChromosomalTheory ofInheritanceand Sexlinkedinheritance

http://www.nature.com/scitable/
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malehasmutation
NormalX chromosome Normal female

Xchromosome withmutation causing hemophilia carrier female Hemophilliac male

femalehasmutation

Sexlinked inheritance
Example:assume therecessive characteristicison theXchromosome XXgenotype= femalephenotype XYgenotype= malephenotype

All daughters= carriers

Allsons= normal

of of sons= daughters daughters normal =normal =carriers

ofsons=
hemophilliac
http://www.accessexcellence.org 55

TheFamousRoyalHemophiliacLine

Greatgrandaughter PresentQueenElizabethII

PrinceCharles PrinceWilliam
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Femalemammalsincluding humansare mosaicduetoX inactivation

Barrbody

or

OneXchromosomeremains active,theother(s)are inactivatedbyaforminghighly condensedBarrbody(red arrow). Xinactivationisrandomduring embryonicdevelopment(1216 days).

Allpinkinactivated

Allblueinactivated

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Femalemammalsincludinghumansaremosaic duetoXinactivation
Acathavingacalicocoat patternisALWAYSfemale. Oneofthegenesresponsible forcoatcolorisontheX chromosome.Therandom distributionoforangeand blackpatchesdemonstratesX inactivation;thewhitepatches areduetoanothergene(s). Humanexample:
Anhydrotic dysplasiaisanXlinkeddiseasethatresultsintheabsenceofsweat glands.NormalfemalesareXXandhavesweatglands,heterozygousfemalesare XX'andhavepatchesofskinwithsweatglandsandpatchesofskinwithout sweatglands.FemalesthatareX'X'donothavesweatglands. 59

Louiseis (probably)a mosaicforeye colorduetoX inactivation

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ChromosomeDynamics: Normalvs.Aberrations
(mutations/abnormalities atthechromosomallevel)
Seeadditionalreadingmaterial: Chromosomalaberration.pdf

61

NormalHomologousRecombination
(occursintestisandovary)

Fromdiploidgermcells

DNA replication

tohaploidsex cells (gametes)

Crossingoverandrecombinationduringmeiosis
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Translocation

63

Insertion

64

Deletion

Duplication

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ArepresentativekaryotypeandaphotographofachildexhibitingCriduchat syndrome (46,5p).Inthekaryotype,thearrowidentifiestheabsenceofa smallpieceoftheshortarmofonememberofthechromosome5homologs.

Affectedindividualshavewideseteyes,asmallheadandjawandare moderatelytoseverelymentallyretardedandveryshort.

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Whenchromosomesegregation fails...
(Only showing meiosis division I)

Normalcase

hasextracopye.g. Chrom.21 Downssyndrome ornocopy

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Nondisjunction duringthefirstandsecondmeioticdivisions.Inbothcases,some ofthegametesthatareformedeithercontaintwomembersofaspecific chromosomeorlackthatchromosome. Afterfertilizationbyagametewitha normalhaploidcontent,monosomic,disomic(normal),ortrisomic zygotesare produced.

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ThekaryotypeofachildwithDownsyndrome.Inthekaryotype, threecopiesofchromosome21arepresent,creatingthe(47,21+) condition.

Characteristicsincludedecreasedmuscletone,asymmetricalskull, slantingeyes,short,flatfacesandmildtomoderatephysical, 69 psychomotorandmentalretardation.

70

Screeningspermcellsorembryostoreducegenetic diseases:Futuredirection?
Thechancesofhavinga babywithasexlinked diseasecanbereducedby sortingspermcells carryingXorY chromosomes(depending onthelocationofthe diseaseallele).

Thiscouldalsobeused forselectingthesex ofthebaby.

Butisthisethical?
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Thisnewtechniqueoffershopeto couplesathighriskofhavingababy withaseriousgeneticcondition. Doctorscancarryoutgenetictestson 'testtube'embryos(i.e.embryos producedusingIVF[invitro fertilization]techniques).Only unaffectedembryosareputbackinto themother'swombtocontinue developing.

Preimplantation GeneticScreening

Screeningembryosisdonefor:
Geneticdiseases(inheritedin Mendelian laws) Susceptibilitytocomplexgenetic diseases HLAtypingforumbilicalcordorbone marrowstemcellstransplantation Gender

Anyethicalissuehere?
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SummaryofKeyConcepts/Points
ClassicalGenetics:inheritanceandtraits
Mendelslawsofheredityexplainhowpaireddiscretefactors(laterknown asgenes)arepassedontooffspring. Genotype&Phenotype,Dominance&recessive(alleles), Heterozygosity &homozygosity.

Whatandwhereisthegeneticmaterial?
Mostlyinnucleus(eukaryoticcells)andusuallyintheformofDNA asdemonstratedbyseveralclassicexperiments.

Celldivision:mitosis&meiosis
Maintenanceofthenumberofchromosomesincellandorganism. Nonsexlinked&sexlinkedinheritance. Punnett square.

Chromosomedynamics:normalvs aberrations
Homologousrecombinationandchromosomeaberrations.
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Chromosome Art

Refer to Supplemental material: DNA ART

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Review:UsingaPunnett square
IntroducedbyReginaldC.Punnett. APunnettsquareisawaytoenvisionallofthe possiblecombinationsthatmaternaland paternalgenescanmakewhencreatinganew organism.

y X

y
FATHER

MOTHER

Punnettsquare

Yy(yellow)andyy(green)

Punnettsquare Y y Yy yy

Y y

YY Yy

Genotypefrequency:YY,Yy,yy

(1:2:1)

YY x yy

all Yy

YY Yy Yy yy

YY Yy Yy yy