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  • EXERCICIO Degenerate - Probes - Exerecicio2013 PDF

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    Oligonucleotide probes are designed based on protein sequence. A 6or 7-aa region can be encoded by y the smallest number of p possible DNA sequences. The region with the least degeneracy for a sequence of 20 bases (20-mer) is indicated by the red bracket.

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    EXERCICIO Degenerate_Probes_Exerecicio2013.pdf

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    Oligonucleotide probes are designed based on protein sequence. A 6or 7-aa region can be encoded by y the smallest number of p possible DNA sequences. The region with the least degeneracy for a sequence of 20 bases (20-mer) is indicated by the red bracket.

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    EXERCICIO Degenerate - Probes - Exerecicio2013 PDF

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    FCiências
    Oligonucleotide probes are designed based on protein sequence. A 6or 7-aa region can be encoded by y the smallest number of p possible DNA sequences. The region with the least degeneracy for a sequence of 20 bases (20-mer) is indicated by the red bracket.

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    Degenerate Probes http://www.ncbi.nlm.nih.

    gov/books/NBK21505/

    From:Section7.3,Identifying,Analyzing,and SequencingClonedDNA MolecularCellBiology.4thedition. Lodish H,Berk A,Zipursky SL,etal. NewYork:W. W H. H Freeman;2000 2000.

    Figure719Designingoligonucleotide probesbasedonproteinsequence Anisolatedproteinisdigestedwithaselectiveproteasesuchastrypsin,whichspecificallycleavespeptidebonds onthecarboxyterminalsideoflysineandarginine residues.

    Theresultingpeptidesareseparated,andseveralarepartiallysequencedfromtheirNterminusbysequential Edman degradation.Thedeterminedsequencesthenareanalyzedtoidentifythe6 or7aaregionthatcanbe encodedby ythesmallestnumberofp possibleDNAsequences. q Becauseofthedegeneracy g yoftheg geneticcode, ,the 12aasequence(lightgreen)shownheretheoreticallycouldbeencodedbyanyoftheDNAtripletsbelowit,with thepossiblealternativebasesatthesamepositionindicated.Forexample,Phe1isencodedbyTTTorTTC;Leu2 isencodedbyoneofsixpossibletriplets(CTT (CTT,CTC CTC,CTA CTA,CTG CTG,TTA TTA,orTTG). TTG) Theregionwiththeleastdegeneracy forasequenceof20bases(20mer)isindicatedbytheredbracket.Thereare48possibleDNAsequencesinthis 20baseregionthatcouldencodethepeptidesequence39.Sincetheactualsequenceofthegeneisunknown, adegenerate20merprobeconsistingofamixtureofallthepossible20baseoligonucleotides isprepared prepared.Ifa cDNA orgenomiclibraryisscreenedwiththisdegenerateprobe,theoneoligonucleotide thatisperfectly complementarytotheactualcodingsequence(blue)willhybridizetoit.

    Exerccio
    Q Qual lseria i acomposio i de d umasonda d degenerada d d de d 20 nucleotdeos paradetectarumfragmentodeDNAcodificante para areferidaprotena? A)umamistura i t de d oligonucleotdeos li l td d 20nt de t quecorrespondesse d aosaminocidoscommaiorgraudedegenerescncia B)umamisturadefragmentosdeDNAde3nt correspondentesa t d oscodes todos d possveis i C)umamisturadefragmentosdeDNAde3nt correspondentesaos oscodes paraaminocidoscommenordegenerescncia D)umoligonucleotdeo de20nt aoqualseadicionaostripletos alternativos E) )umamisturadeoligonucleotdeos g de20nt q quecorrespondesse p aosaminocidoscommenorgraudedegenerescncia

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