BCH 5413- EXAM 1 Questions And Answers

BCH 5413- EXAM 1 Questions And
Answers
2ndrgenerationrplasmidr-rCORRECTrrANSWER-pUC19r(oric,rlacZ,rAp^R)

ABrfragmentsrselectedrusingr***r-rCORRECTrrANSWER-Avidin-biotinrpurification

Apyraser-rCORRECTrrANSWER-
clearsroutrintermediatesrandrsubstratesrafterrfirstrreactionr(454)

autoradiographyr-rCORRECTrrANSWER-
israrmeansrofrdetectionrusingrradioactivelyrlabeledrDNA,rwhenrtheserhigh-
energyrelectronsrstrikertherscreen,rtheyrcauserfluorescence,rwhichrisrdetectedrbyrtherfilm.

BamH1rrecognitionrsequencer-rCORRECTrrANSWER-Gr↓rGrArTrCrC

Bridgeramplificationr-rCORRECTrrANSWER-
addrunlabeledrnucleotidesrandrenzymertorinitialrsolid-phaserbridgeramplification

cDNArproberstartingrpointsr-rCORRECTrrANSWER-
1.rsamerspecies:r65rdegreerinr6XSCCrwashr65rdegreerinr1/10rSSC)
2.rdifferentrrelatedrspecies:r60r6rxrSCC,rwashr65r1xrSCC
3.rdistantrspecies:r60rdegreerinr6xSCC,rwashr55-60rdegreerinr1xrSCC

Cellrembryologyr-rCORRECTrrANSWER-4rcell--
>rMorularatr2.5rdaysrwhichrisrundifferentiatedr--
>rBlastocyteratr3.5rdaysrwhichrhasr2rcellrtypesr(Traphectodermrandrinnerrcellrmass)
**innerrcellrmassrbecomesrembyro

Charonr4r-rCORRECTrrANSWER-Vectorraccommmodatesrinsertsrbetweenr12-20kB

Chimericrmouser-rCORRECTrrANSWER-
igurer5.43rMakingrarknockoutrmouse:rStager2,rplacingrthe
interruptedrgenerinrtheranimal.r(1)rInjectrthercellsrwithrthe
rinterruptedrgener(seerFigurer5.42)rintorarblastocyst-stagerembryorfrom

, blackrparentrmice.r(2)rTransplantrthisrmixedrembryortortheruterusrofra
surrogatermother.r(3)rThersurrogatermotherrgivesrbirthrtorarchimeric
mouse,rwhichronercanridentifyrbyritsrblackrandrbrownrcoat.r(Recall
thatrtheralteredrcellsrcamerfromranragoutir[brown]rmouse,randrthey
wererplacedrintoranrembryorfromrarblackrmouse.)r(4)rAllowrthe
chimericrmouser(armale)rtormature.r(5)rMateritrwithrarwild-typerblack
female.rDiscardranyrblackroffspring,rwhichrmustrhaverderivedrfromrthe
wild-typerblastocyst;ronlyrbrownrmicercouldrhaverderivedrfromrthe
transplantedrcells.r(6)rSelectrarbrownrbrotherrandrsisterrpair,rbothrof
whichrshowrevidencerofranrinterruptedrtargetrgener(byrSouthernrblot
analysis),randrmaterthem.rAgain,rexaminertherDNArofrtherbrown
progenyrbyrSouthernrblotting.rThisrtime,roneranimalrthatris
homozygousrforrtherinterruptedrtargetrgenerisrfound.rThisrisrthe
knockoutrmouse.rNowrobserverthisranimalrtordeterminerthereffectsrof
knockingroutrthertargetrgene.

competentr-rCORRECTrrANSWER-
TreatedrwithrCaCl2rtorweakenrmembranerandrincreaserlikelihoodrofrbeingrtakenruprbyrbacte
rialrcell

ComponentsrofrPlasmidr-rCORRECTrrANSWER-1.rOriCr(originrofrreplication)
2.rSelectablermarkerreitherrApr^Rr(ampicillinrresistance)rorrTcr^Rr(tetracyclinerresistance)
3.rpolylinkerr(MSC)
4.rLacrZ

Cross-reactionsr-rCORRECTrrANSWER-
Couldronerofryourrreagentsrberdetectingrsomethingrelserasrwell?
userBLASTr(NCIBrwebsite)

CT:r-rCORRECTrrANSWER-
whenrthercyclernumberrcrossesrarthreshold,rlowerrCTrmeansrmorerproductrisrpresent

Digestrwithr__r-rCORRECTrrANSWER-EcoR1

Dimethylsulfater-rCORRECTrrANSWER-modifiesrG

DNArFootprintingr-rCORRECTrrANSWER-usedrtoranalyzersequence-specificrDNA-
proteinrinteractions

•DNArfootprintingroccursrwhenrdigestionrofrarspecificrregionrofrDNArisrpreventedrbyrbindingr
ofrarprotein.rTherprotectionrcanrbereitherrtordigestionrbyrDNases,rorrtorchemicalrmodification
.rThersecondrprocessrgivesrgreaterrresolution.
r

***Therfootprintrrepresentsrtherregionrof
DNArprotectedrbyrtherprotein,randrthereforertellsrwhere
therproteinrbinds