BLAST nucleotideo (BLASTn)

O programa blastn é usado quanto temos uma sequência de DNA (query) e queremos buscar a sequência de DNA (subject) do banco de dados similar a nossa sequência, são chamados de alinhamentos nucleotídeo-nucleotídeo. As comparações nucleotídeo-nucleotídeo tem duas variantes dentro da família BLAST, o programa blastn e o programa MegaBLAST.

O programa blastn é usado para fazer buscas e alinhamentos de segmentos de DNA mais gerais, usando regiões de genomas onde encontramos misturas de regiões codificantes e não codificantes, comparaçoes de rRNA, tRNA e mRNA.

O MegaBLAST faz a comparação 10 vezes mais rápida que o blastn, mas ele busca sequências que são extremamente similares, permitindo mapeamento de transcritos no genoma de 3 bilhões de bases em segundos, e útil para processamento de um grande conjunto de sequências. Uma variante do MegaBLAST é o “discontiguous MegaBLAST”. O discontiguous MegaBLAST é uma programa que que não exisge o pareamento a terceira base em uma comapração de sequencias codificadores de proteínas, como ocorre com as wobble base position dos códons durante a tradução. Essa ferramente permite a busca de sequencias similares entre espécies relacionadas que apresentam diferenças na terceira posição dos codons, portanto, não seriam encontradas pelo MegaBLAST.

Problema 1.

Figura 1. Resultado gráfico do blastn. Existem 5 sequências alinhando com o primer forward e seis sequências alinhando com o primer reverse.

 

Figura 2. Alinhamento dos primers forward e reverse com a sequência subject. O primer forward alinhou com a sequência subject na fita reversa (Plus/Minus) e o primer reverse alinhou com a sequência subject na fita direta (Plus/Plus).

O resultado dessa busca retornou 11 entradas no GenBank (BLAST hits) que alinham em ambos os primires foward e reverse em difertentes posições indicados pelas barras mais grossas com um gap entre elas indicado pela linha em cinza ligando duas barras grossas.

Existe uma entrada no GenBank que alinha com somente o primer reverse. Um alinhamento do par de primer é mostrado na figura 2. O primer foward alinha na fita reverse (indicado pelo Strand Plus/Minus) no nuleodideo 41513018..41513000. O primer reverso alinha na fita foward (indicado pela Strand Plus/Plus) na posição 41512795..41512795. Então esses dois primers irão amplificar a sequência da posição 41512795..41513018 da entrada NC_018928.2. Recupere a seqêencia do Entrez clicando sobre ela. A anotação da sequência mostra que ela cobre uma região do exon e a parte upstream do gene BRCA1.

Exercicío

Vamos agora ver como o BLAST pode ser útil para estudar polimorfismo. Vamos usar o MegaBLAST para buscar sequências similares e observar esses polimorfismos em um alinhamento múltiplo. Hermankova, M. et al. (2001) estudou resistência a droga em HIV-1 avaliando pacientes recebendo terapia. Vamos identificar os SNPs em sequências HIV-1.

Façam a busca usando a seqência abaixo

>DNA1
ATGACCTCAAATCACTCTTTGGCAACGACCCCTCGTCACAATAAAGATAGGGGGGCAACTAAAGGAAGCTCTATTAGATACAGGAGCAGATGATACAGTATTAGAAGAAATGGAGTTGCCAGGAAGATGGAAACCAAAAATGATAGGGGGAATTGGAGGTTTTATCAAAGTAAGACAGTATGATCAGATACCCATAGAAATCTGTGGACATAGAGCTATAGGTACAGTATTAGTAGGACCTACACCTGTCAACATAATTGGAAGAAATCTGTTGACTCAGCTTGGTTGCACTTTAAATTTTCCCATAAGTCCTATTGAAACTGTACCAGTAAAATTAAAGCCAGGAATGGATGGCCCAAAAGTTAAACAATGGCCATTGACAGAAGAAAAAATAAAAGCATTAGTAGAAATTTGTACAGAAATGGAAAAGGAAGGGAAAATTTCAAAAATTGGGCCTGAAAATCCATACAATACTCCAGTATTTGCCATAAAGAAAAAAGACAGTACTAAATGGAGAAAATTAGTAGATTTCAGAGAACTTAATAAGAAAACTCAAGACTTCTGGGAAGTTCAATTAGGAATACCACATCCCGCAGGGTTAAAAAAGAAAAAATCAGTAACAGTACTGGATGTGGGTGATGCATATTTTTCAGTTCCCTTAGATAAAGACTTCAGGAAGTACACTGCATTTACCATACCTAGTATAAACAATGAGACACCAGGGATTAGATATCAGTACAATGTGCTTCCACAGGGATGGAAAGGATCACCAGCAATATTCCAAAGTAGTATGACAAAAATTTTAGAGCCTTTTAGAGAACAAAATCCAGAAATAGTTATCTATCAATACATGGATGATTTGTATGTAGGATCTGACTTAGAACTAGGGCAGCATAGAGCAAAAATAGAGGAACTGAGACGACATCTGTTGAGGTGGGGATTTACCACACCAGACAAAAAACATCAGAAAGAGCCTCCATTCCTTTGGA

deletion
ATTTCAAAAATTGGGCCTGAAAATCCATACAATACTCCAGTATTTGCCATAAAGAAAAAAGACAGTACTA
AATGGAGAAAATTAGTAGATTTCAGAGAACTTAATAAGAAAACGCAAGACTTTTGGGAAGTCCAATTAGG
AATACCACACCCTGCAGGGTTAAAAAAGAAAAAATCAGTAACAGTACTGGATGTGGGTGATGCATATTTT
TCAGTTCCCTTAGATAAAGACTTCAGGAAGTATACTGCATTTACCATACCTAGTACAAACAATGAGACAC
CAGGGATTAGATATCAGTACAATGTGCTTCCACAGGGATGGAAAGGATCACCAGCAATATTCCAGTGTAG
CATGACAAAAATCTTAGAGCCTTTTAGAAAACAAAATCCAGACCTAGTTATCTATCAATACATGGATGAT
TTGTATGTAGGATCTGACTTAGAACTAGGGCAGCATAGAACAAAAATAGAGGAACTGAGACAACATCTGT
TGAAGTGGGGGTTTACTACACCAGACAAAAAACATCAGAAAGAACCTCCATTCCTTTGGATGGGTTATGA
ACTCCATCCTGATAAATGGACAGTACAGCCTATAGTGCTGCCAGAAAAAGACAGCTGGACTGTCAATGAC
ATACAGAAGTTAGTGGGAAAATTGAATTGGGCAAGTCAGATTTATGCAGGGATCAAAGTAAAGCAATTAT
GTAAACTCCTTAGGGGAACTAAAGCACTAACAGAAGTAATACCACTAACAGAAGAAGCAGAACTAGAGCT
GGCAGAAAATAGGGAAATTCTAAAAGAACCAGTACATGGGGTGTATTATGACCCAACAAAAGACCTAATA
GCAGAATTACAGAAGCAGGGGCAAGGCCAATGGACCTATCAAATTTATCAAGAGCCATTTAAAAATCTGA
AAACAGGAAAATATGCAAGAATGAGGGGTGCCCACACTAATGATGTAAAACAGTTAACAGAGGCAGTGCA
AAAAATAGCCACAGAAAGCATAGTAATATGGGGAAAAATTCCTAAATTTAAGCTACCCATACAAAAAGAA
ACATGGGAAACATGGTGGACAGAGTATTGGCAAGCCACCTGGATTCCTGAGTGGGAGTTTGTTAATACCC
CTCCCTTAGTGAAATTATGGTACCAGTTAGAGAAAGAACCCATAGTAGGAGCAGAAACTTTCTATGTAGA
TGGGGCAGCTAACAGGGAAACTAAATTAGGAAAAGCAGGATATGTTACTGACAGAGGAAGACAAAAAGTT
GTCTCCCTAACTGACACAACAAATCAGAAGACTGAGTTACAAGCAATTCATCTAGCTTTGCAGGATTCGG
GATTAGAAGTAAACATAGTAACAGACTCACAATATGCATTAGGAATCATTCAAGCACAACCAGATAAAAG
TGAATCAGAGTTAGTCAGTCAAATAATAGAGCAGTTAATAAAAAAGGAAAAGGTCTACCTGACATGGGTA
TTAGCGATTATCTGGGTCGATCTGCGGAGCCTGTGCCTCTTCAGCTACCACCGCTTGAGAGACTTACTCT
TGATTGTAGCGAGGATTGTGGAACTTCTGGGACACAGGGGGTGGGAGATCCTCAAATATTGGGGGAATCT
CCTACAATATTGGAGTCAGGAACTAAAGAATAGTGCTGTTAGCTTGCTCAACGCCACAGCCATAGCAGTA
GCTGAGGGGACAGATAGAATTATAGAAGTAGTACAAAGAGCTGGTAGGGCTATTCTCCACATACCTGTAA
GAATAAGACAGGGCTTGGAAAGGGCTTTGCTATAAGATGGGTGGCAAGTGGTCAAAAAGTAGTATGGTTG
GATGGCCTACTATAAGGGAAAGAATGAAACGAGCTGAGCCAGCAGCAGTTGGGGTGGGAGCAGCATCTCG
AGACCTGGATAAACATGGAGCAATCACAAGTAGCAATACAGCAGCTAACAATGCTGATTGTGCCTGGCTA
GAAGCACAAGAGGAGGAGGTGGGTTTTCCAGTCAGACCTCAGGTACCTCTAAGACCAATGACTTACAAGG
CAGCTGTAGATCTTAGCCACTTTTTAAAAGAAAAGGGGGGACTGGAAGGGCTAATTTACTCCCAAAAAAG
ACAAGATATCCTTGATTTATGGGTCTACCACACACAAGGCTACTTCCCTGATTGGCAGAACTACACACCA

Vamos mudar a “Formatting options” para observar o SNP.

Vamos alterar a opção:“Alignment View”

Selecionem: “Flat query-anchored with dots for identities”

Clicar em “Reformat”

Figura 3. Alterando o formato do resultado do blastn para mostrar um alinhamento com pontos para as posições idênticas a query e letras para as bases diferentes da query.

Procurem onde estão os polimorfismos. Descrevam os SNPs da seguinte forma.

Exemplo: Posição 50 bases C e T, representem 50C/T.

Descrevam os SNPs para as três sequências abaixo

SEQUENCIA1

SEQUENCIA2

SEQUENCIA3

>gi|588282786|ref|NM_002111.7| Homo sapiens huntingtin (HTT), mRNA

TTGCTGTGTGAGGCAGAACCTGCGGGGGCAGGGGCGGGCTGGTTCCCTGGCCAGCCATTGGCAGAGTCCG
CAGGCTAGGGCTGTCAATCATGCTGGCCGGCGTGGCCCCGCCTCCGCCGGCGCGGCCCCGCCTCCGCCGG
CGCAGCGTCTGGGACGCAAGGCGCCGTGGGGGCTGCCGGGACGGGTCCAAGATGGACGGCCGCTCAGGTT
CTGCTTTTACCTGCGGCCCAGAGCCCCATTCATTGCCCCGGTGCTGAGCGGCGCCGCGAGTCGGCCCGAG
GCCTCCGGGGACTGCCGTGCCGGGCGGGAGACCGCCATGGCGACCCTGGAAAAGCTGATGAAGGCCTTCG
AGTCCCTCAAGTCCTTCCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCAGCA
GCAGCAGCAGCAACAGCCGCCACCGCCGCCGCCGCCGCCGCCGCCTCCTCAGCTTCCTCAGCCGCCGCCG
CAGGCACAGCCGCTGCTGCCTCAGCCGCAGCCGCCCCCGCCGCCGCCCCCGCCGCCACCCGGCCCGGCTG
TGGCTGAGGAGCCGCTGCACCGACCAAAGAAAGAACTTTCAGCTACCAAGAAAGACCGTGTGAATCATTG
TCTGACAATATGTGAAAACATAGTGGCACAGTCTGTCAGAAATTCTCCAGAATTTCAGAAACTTCTGGGC
ATCGCTATGGAACTTTTTCTGCTGTGCAGTGATGACGCAGAGTCAGATGTCAGGATGGTGGCTGACGAAT
GCCTCAACAAAGTTATCAAAGCTTTGATGGATTCTAATCTTCCAAGGTTACAGCTCGAGCTCTATAAGGA
AATTAAAAAGAATGGTGCCCCTCGGAGTTTGCGTGCTGCCCTGTGGAGGTTTGCTGAGCTGGCTCACCTG
GTTCGGCCTCAGAAATGCAGGCCTTACCTGGTGAACCTTCTGCCGTGCCTGACTCGAACAAGCAAGAGAC
CCGAAGAATCAGTCCAGGAGACCTTGGCTGCAGCTGTTCCCAAAATTATGGCTTCTTTTGGCAATTTTGC
AAATGACAATGAAATTAAGGTTTTGTTAAAGGCCTTCATAGCGAACCTGAAGTCAAGCTCCCCCACCATT
CGGCGGACAGCGGCTGGATCAGCAGTGAGCATCTGCCAGCACTCAAGAAGGACACAATATTTCTATAGTT
GGCTACTAAATGTGCTCTTAGGCTTACTCGTTCCTGTCGAGGATGAACACTCCACTCTGCTGATTCTTGG
CGTGCTGCTCACCCTGAGGTATTTGGTGCCCTTGCTGCAGCAGCAGGTCAAGGACACAAGCCTGAAAGGC
AGCTTCGGAGTGACAAGGAAAGAAATGGAAGTCTCTCCTTCTGCAGAGCAGCTTGTCCAGGTTTATGAAC
TGACGTTACATCATACACAGCACCAAGACCACAATGTTGTGACCGGAGCCCTGGAGCTGTTGCAGCAGCT
CTTCAGAACGCCTCCACCCGAGCTTCTGCAAACCCTGACCGCAGTCGGGGGCATTGGGCAGCTCACCGCT
GCTAAGGAGGAGTCTGGTGGCCGAAGCCGTAGTGGGAGTATTGTGGAACTTATAGCTGGAGGGGGTTCCT
CATGCAGCCCTGTCCTTTCAAGAAAACAAAAAGGCAAAGTGCTCTTAGGAGAAGAAGAAGCCTTGGAGGA
TGACTCTGAATCGAGATCGGATGTCAGCAGCTCTGCCTTAACAGCCTCAGTGAAGGATGAGATCAGTGGA
GAGCTGGCTGCTTCTTCAGGGGTTTCCACTCCAGGGTCAGCAGGTCATGACATCATCACAGAACAGCCAC
GGTCACAGCACACACTGCAGGCGGACTCAGTGGATCTGGCCAGCTGTGACTTGACAAGCTCTGCCACTGA
TGGGGATGAGGAGGATATCTTGAGCCACAGCTCCAGCCAGGTCAGCGCCGTCCCATCTGACCCTGCCATG
GACCTGAATGATGGGACCCAGGCCTCGTCGCCCATCAGCGACAGCTCCCAGACCACCACCGAAGGGCCTG
ATTCAGCTGTTACCCCTTCAGACAGTTCTGAAATTGTGTTAGACGGTACCGACAACCAGTATTTGGGCCT
GCAGATTGGACAGCCCCAGGATGAAGATGAGGAAGCCACAGGTATTCTTCCTGATGAAGCCTCGGAGGCC
TTCAGGAACTCTTCCATGGCCCTTCAACAGGCACATTTATTGAAAAACATGAGTCACTGCAGGCAGCCTT
CTGACAGCAGTGTTGATAAATTTGTGTTGAGAGATGAAGCTACTGAACCGGGTGATCAAGAAAACAAGCC
TTGCCGCATCAAAGGTGACATTGGACAGTCCACTGATGATGACTCTGCACCTCTTGTCCATTGTGTCCGC
CTTTTATCTGCTTCGTTTTTGCTAACAGGGGGAAAAAATGTGCTGGTTCCGGACAGGGATGTGAGGGTCA
GCGTGAAGGCCCTGGCCCTCAGCTGTGTGGGAGCAGCTGTGGCCCTCCACCCGGAATCTTTCTTCAGCAA
ACTCTATAAAGTTCCTCTTGACACCACGGAATACCCTGAGGAACAGTATGTCTCAGACATCTTGAACTAC
ATCGATCATGGAGACCCACAGGTTCGAGGAGCCACTGCCATTCTCTGTGGGACCCTCATCTGCTCCATCC
TCAGCAGGTCCCGCTTCCACGTGGGAGATTGGATGGGCACCATTAGAACCCTCACAGGAAATACATTTTC
TTTGGCGGATTGCATTCCTTTGCTGCGGAAAACACTGAAGGATGAGTCTTCTGTTACTTGCAAGTTAGCT
TGTACAGCTGTGAGGAACTGTGTCATGAGTCTCTGCAGCAGCAGCTACAGTGAGTTAGGACTGCAGCTGA
TCATCGATGTGCTGACTCTGAGGAACAGTTCCTATTGGCTGGTGAGGACAGAGCTTCTGGAAACCCTTGC
AGAGATTGACTTCAGGCTGGTGAGCTTTTTGGAGGCAAAAGCAGAAAACTTACACAGAGGGGCTCATCAT
TATACAGGGCTTTTAAAACTGCAAGAACGAGTGCTCAATAATGTTGTCATCCATTTGCTTGGAGATGAAG
ACCCCAGGGTGCGACATGTTGCCGCAGCATCACTAATTAGGCTTGTCCCAAAGCTGTTTTATAAATGTGA
CCAAGGACAAGCTGATCCAGTAGTGGCCGTGGCAAGAGATCAAAGCAGTGTTTACCTGAAACTTCTCATG
CATGAGACGCAGCCTCCATCTCATTTCTCCGTCAGCACAATAACCAGAATATATAGAGGCTATAACCTAC
TACCAAGCATAACAGACGTCACTATGGAAAATAACCTTTCAAGAGTTATTGCAGCAGTTTCTCATGAACT
AATCACATCAACCACCAGAGCACTCACATTTGGATGCTGTGAAGCTTTGTGTCTTCTTTCCACTGCCTTC
CCAGTTTGCATTTGGAGTTTAGGTTGGCACTGTGGAGTGCCTCCACTGAGTGCCTCAGATGAGTCTAGGA
AGAGCTGTACCGTTGGGATGGCCACAATGATTCTGACCCTGCTCTCGTCAGCTTGGTTCCCATTGGATCT
CTCAGCCCATCAAGATGCTTTGATTTTGGCCGGAAACTTGCTTGCAGCCAGTGCTCCCAAATCTCTGAGA
AGTTCATGGGCCTCTGAAGAAGAAGCCAACCCAGCAGCCACCAAGCAAGAGGAGGTCTGGCCAGCCCTGG
GGGACCGGGCCCTGGTGCCCATGGTGGAGCAGCTCTTCTCTCACCTGCTGAAGGTGATTAACATTTGTGC
CCACGTCCTGGATGACGTGGCTCCTGGACCCGCAATAAAGGCAGCCTTGCCTTCTCTAACAAACCCCCCT
TCTCTAAGTCCCATCCGACGAAAGGGGAAGGAGAAAGAACCAGGAGAACAAGCATCTGTACCGTTGAGTC
CCAAGAAAGGCAGTGAGGCCAGTGCAGCTTCTAGACAATCTGATACCTCAGGTCCTGTTACAACAAGTAA
ATCCTCATCACTGGGGAGTTTCTATCATCTTCCTTCATACCTCAAACTGCATGATGTCCTGAAAGCTACA
CACGCTAACTACAAGGTCACGCTGGATCTTCAGAACAGCACGGAAAAGTTTGGAGGGTTTCTCCGCTCAG
CCTTGGATGTTCTTTCTCAGATACTAGAGCTGGCCACACTGCAGGACATTGGGAAGTGTGTTGAAGAGAT
CCTAGGATACCTGAAATCCTGCTTTAGTCGAGAACCAATGATGGCAACTGTTTGTGTTCAACAATTGTTG
AAGACTCTCTTTGGCACAAACTTGGCCTCCCAGTTTGATGGCTTATCTTCCAACCCCAGCAAGTCACAAG
GCCGAGCACAGCGCCTTGGCTCCTCCAGTGTGAGGCCAGGCTTGTACCACTACTGCTTCATGGCCCCGTA
CACCCACTTCACCCAGGCCCTCGCTGACGCCAGCCTGAGGAACATGGTGCAGGCGGAGCAGGAGAACGAC
ACCTCGGGATGGTTTGATGTCCTCCAGAAAGTGTCTACCCAGTTGAAGACAAACCTCACGAGTGTCACAA
AGAACCGTGCAGATAAGAATGCTATTCATAATCACATTCGTTTGTTTGAACCTCTTGTTATAAAAGCTTT
AAAACAGTACACGACTACAACATGTGTGCAGTTACAGAAGCAGGTTTTAGATTTGCTGGCGCAGCTGGTT
CAGTTACGGGTTAATTACTGTCTTCTGGATTCAGATCAGGTGTTTATTGGCTTTGTATTGAAACAGTTTG
AATACATTGAAGTGGGCCAGTTCAGGGAATCAGAGGCAATCATTCCAAACATCTTTTTCTTCTTGGTATT
ACTATCTTATGAACGCTATCATTCAAAACAGATCATTGGAATTCCTAAAATCATTCAGCTCTGTGATGGC
ATCATGGCCAGTGGAAGGAAGGCTGTGACACATGCCATACCGGCTCTGCAGCCCATAGTCCACGACCTCT
TTGTATTAAGAGGAACAAATAAAGCTGATGCAGGAAAAGAGCTTGAAACCCAAAAAGAGGTGGTGGTGTC
AATGTTACTGAGACTCATCCAGTACCATCAGGTGTTGGAGATGTTCATTCTTGTCCTGCAGCAGTGCCAC
AAGGAGAATGAAGACAAGTGGAAGCGACTGTCTCGACAGATAGCTGACATCATCCTCCCAATGTTAGCCA
AACAGCAGATGCACATTGACTCTCATGAAGCCCTTGGAGTGTTAAATACATTATTTGAGATTTTGGCCCC
TTCCTCCCTCCGTCCGGTAGACATGCTTTTACGGAGTATGTTCGTCACTCCAAACACAATGGCGTCCGTG
AGCACTGTTCAACTGTGGATATCGGGAATTCTGGCCATTTTGAGGGTTCTGATTTCCCAGTCAACTGAAG
ATATTGTTCTTTCTCGTATTCAGGAGCTCTCCTTCTCTCCGTATTTAATCTCCTGTACAGTAATTAATAG
GTTAAGAGATGGGGACAGTACTTCAACGCTAGAAGAACACAGTGAAGGGAAACAAATAAAGAATTTGCCA
GAAGAAACATTTTCAAGGTTTCTATTACAACTGGTTGGTATTCTTTTAGAAGACATTGTTACAAAACAGC
TGAAGGTGGAAATGAGTGAGCAGCAACATACTTTCTATTGCCAGGAACTAGGCACACTGCTAATGTGTCT
GATCCACATCTTCAAGTCTGGAATGTTCCGGAGAATCACAGCAGCTGCCACTAGGCTGTTCCGCAGTGAT
GGCTGTGGCGGCAGTTTCTACACCCTGGACAGCTTGAACTTGCGGGCTCGTTCCATGATCACCACCCACC
CGGCCCTGGTGCTGCTCTGGTGTCAGATACTGCTGCTTGTCAACCACACCGACTACCGCTGGTGGGCAGA
AGTGCAGCAGACCCCGAAAAGACACAGTCTGTCCAGCACAAAGTTACTTAGTCCCCAGATGTCTGGAGAA
GAGGAGGATTCTGACTTGGCAGCCAAACTTGGAATGTGCAATAGAGAAATAGTACGAAGAGGGGCTCTCA
TTCTCTTCTGTGATTATGTCTGTCAGAACCTCCATGACTCCGAGCACTTAACGTGGCTCATTGTAAATCA
CATTCAAGATCTGATCAGCCTTTCCCACGAGCCTCCAGTACAGGACTTCATCAGTGCCGTTCATCGGAAC
TCTGCTGCCAGCGGCCTGTTCATCCAGGCAATTCAGTCTCGTTGTGAAAACCTTTCAACTCCAACCATGC
TGAAGAAAACTCTTCAGTGCTTGGAGGGGATCCATCTCAGCCAGTCGGGAGCTGTGCTCACGCTGTATGT
GGACAGGCTTCTGTGCACCCCTTTCCGTGTGCTGGCTCGCATGGTCGACATCCTTGCTTGTCGCCGGGTA
GAAATGCTTCTGGCTGCAAATTTACAGAGCAGCATGGCCCAGTTGCCAATGGAAGAACTCAACAGAATCC
AGGAATACCTTCAGAGCAGCGGGCTCGCTCAGAGACACCAAAGGCTCTATTCCCTGCTGGACAGGTTTCG
TCTCTCCACCATGCAAGACTCACTTAGTCCCTCTCCTCCAGTCTCTTCCCACCCGCTGGACGGGGATGGG
CACGTGTCACTGGAAACAGTGAGTCCGGACAAAGACTGGTACGTTCATCTTGTCAAATCCCAGTGTTGGA
CCAGGTCAGATTCTGCACTGCTGGAAGGTGCAGAGCTGGTGAATCGGATTCCTGCTGAAGATATGAATGC
CTTCATGATGAACTCGGAGTTCAACCTAAGCCTGCTAGCTCCATGCTTAAGCCTAGGGATGAGTGAAATT
TCTGGTGGCCAGAAGAGTGCCCTTTTTGAAGCAGCCCGTGAGGTGACTCTGGCCCGTGTGAGCGGCACCG
TGCAGCAGCTCCCTGCTGTCCATCATGTCTTCCAGCCCGAGCTGCCTGCAGAGCCGGCGGCCTACTGGAG
CAAGTTGAATGATCTGTTTGGGGATGCTGCACTGTATCAGTCCCTGCCCACTCTGGCCCGGGCCCTGGCA
CAGTACCTGGTGGTGGTCTCCAAACTGCCCAGTCATTTGCACCTTCCTCCTGAGAAAGAGAAGGACATTG
TGAAATTCGTGGTGGCAACCCTTGAGGCCCTGTCCTGGCATTTGATCCATGAGCAGATCCCGCTGAGTCT
GGATCTCCAGGCAGGGCTGGACTGCTGCTGCCTGGCCCTGCAGCTGCCTGGCCTCTGGAGCGTGGTCTCC
TCCACAGAGTTTGTGACCCACGCCTGCTCCCTCATCTACTGTGTGCACTTCATCCTGGAGGCCGTTGCAG
TGCAGCCTGGAGAGCAGCTTCTTAGTCCAGAAAGAAGGACAAATACCCCAAAAGCCATCAGCGAGGAGGA
GGAGGAAGTAGATCCAAACACACAGAATCCTAAGTATATCACTGCAGCCTGTGAGATGGTGGCAGAAATG
GTGGAGTCTCTGCAGTCGGTGTTGGCCTTGGGTCATAAAAGGAATAGCGGCGTGCCGGCGTTTCTCACGC
CATTGCTAAGGAACATCATCATCAGCCTGGCCCGCCTGCCCCTTGTCAACAGCTACACACGTGTGCCCCC
ACTGGTGTGGAAGCTTGGATGGTCACCCAAACCGGGAGGGGATTTTGGCACAGCATTCCCTGAGATCCCC
GTGGAGTTCCTCCAGGAAAAGGAAGTCTTTAAGGAGTTCATCTACCGCATCAACACACTAGGCTGGACCA
GTCGTACTCAGTTTGAAGAAACTTGGGCCACCCTCCTTGGTGTCCTGGTGACGCAGCCCCTCGTGATGGA
GCAGGAGGAGAGCCCACCAGAAGAAGACACAGAGAGGACCCAGATCAACGTCCTGGCCGTGCAGGCCATC
ACCTCACTGGTGCTCAGTGCAATGACTGTGCCTGTGGCCGGCAACCCAGCTGTAAGCTGCTTGGAGCAGC
AGCCCCGGAACAAGCCTCTGAAAGCTCTCGACACCAGGTTTGGGAGGAAGCTGAGCATTATCAGAGGGAT
TGTGGAGCAAGAGATTCAAGCAATGGTTTCAAAGAGAGAGAATATTGCCACCCATCATTTATATCAGGCA
TGGGATCCTGTCCCTTCTCTGTCTCCGGCTACTACAGGTGCCCTCATCAGCCACGAGAAGCTGCTGCTAC
AGATCAACCCCGAGCGGGAGCTGGGGAGCATGAGCTACAAACTCGGCCAGGTGTCCATACACTCCGTGTG
GCTGGGGAACAGCATCACACCCCTGAGGGAGGAGGAATGGGACGAGGAAGAGGAGGAGGAGGCCGACGCC
CCTGCACCTTCGTCACCACCCACGTCTCCAGTCAACTCCAGGAAACACCGGGCTGGAGTTGACATCCACT
CCTGTTCGCAGTTTTTGCTTGAGTTGTACAGCCGCTGGATCCTGCCGTCCAGCTCAGCCAGGAGGACCCC
GGCCATCCTGATCAGTGAGGTGGTCAGATCCCTTCTAGTGGTCTCAGACTTGTTCACCGAGCGCAACCAG
TTTGAGCTGATGTATGTGACGCTGACAGAACTGCGAAGGGTGCACCCTTCAGAAGACGAGATCCTCGCTC
AGTACCTGGTGCCTGCCACCTGCAAGGCAGCTGCCGTCCTTGGGATGGACAAGGCCGTGGCGGAGCCTGT
CAGCCGCCTGCTGGAGAGCACGCTCAGGAGCAGCCACCTGCCCAGCAGGGTTGGAGCCCTGCACGGCGTC
CTCTATGTGCTGGAGTGCGACCTGCTGGACGACACTGCCAAGCAGCTCATCCCGGTCATCAGCGACTATC
TCCTCTCCAACCTGAAAGGGATCGCCCACTGCGTGAACATTCACAGCCAGCAGCACGTACTGGTCATGTG
TGCCACTGCGTTTTACCTCATTGAGAACTATCCTCTGGACGTAGGGCCGGAATTTTCAGCATCAATAATA
CAGATGTGTGGGGTGATGCTGTCTGGAAGTGAGGAGTCCACCCCCTCCATCATTTACCACTGTGCCCTCA
GAGGCCTGGAGCGCCTCCTGCTCTCTGAGCAGCTCTCCCGCCTGGATGCAGAATCGCTGGTCAAGCTGAG
TGTGGACAGAGTGAACGTGCACAGCCCGCACCGGGCCATGGCGGCTCTGGGCCTGATGCTCACCTGCATG
TACACAGGAAAGGAGAAAGTCAGTCCGGGTAGAACTTCAGACCCTAATCCTGCAGCCCCCGACAGCGAGT
CAGTGATTGTTGCTATGGAGCGGGTATCTGTTCTTTTTGATAGGATCAGGAAAGGCTTTCCTTGTGAAGC
CAGAGTGGTGGCCAGGATCCTGCCCCAGTTTCTAGACGACTTCTTCCCACCCCAGGACATCATGAACAAA
GTCATCGGAGAGTTTCTGTCCAACCAGCAGCCATACCCCCAGTTCATGGCCACCGTGGTGTATAAGGTGT
TTCAGACTCTGCACAGCACCGGGCAGTCGTCCATGGTCCGGGACTGGGTCATGCTGTCCCTCTCCAACTT
CACGCAGAGGGCCCCGGTCGCCATGGCCACGTGGAGCCTCTCCTGCTTCTTTGTCAGCGCGTCCACCAGC
CCGTGGGTCGCGGCGATCCTCCCACATGTCATCAGCAGGATGGGCAAGCTGGAGCAGGTGGACGTGAACC
TTTTCTGCCTGGTCGCCACAGACTTCTACAGACACCAGATAGAGGAGGAGCTCGACCGCAGGGCCTTCCA
GTCTGTGCTTGAGGTGGTTGCAGCCCCAGGAAGCCCATATCACCGGCTGCTGACTTGTTTACGAAATGTC
CACAAGGTCACCACCTGCTGAGCGCCATGGTGGGAGAGACTGTGAGGCGGCAGCTGGGGCCGGAGCCTTT
GGAAGTCTGCGCCCTTGTGCCCTGCCTCCACCGAGCCAGCTTGGTCCCTATGGGCTTCCGCACATGCCGC
GGGCGGCCAGGCAACGTGCGTGTCTCTGCCATGTGGCAGAAGTGCTCTTTGTGGCAGTGGCCAGGCAGGG
AGTGTCTGCAGTCCTGGTGGGGCTGAGCCTGAGGCCTTCCAGAAAGCAGGAGCAGCTGTGCTGCACCCCA
TGTGGGTGACCAGGTCCTTTCTCCTGATAGTCACCTGCTGGTTGTTGCCAGGTTGCAGCTGCTCTTGCAT
CTGGGCCAGAAGTCCTCCCTCCTGCAGGCTGGCTGTTGGCCCCTCTGCTGTCCTGCAGTAGAAGGTGCCG
TGAGCAGGCTTTGGGAACACTGGCCTGGGTCTCCCTGGTGGGGTGTGCATGCCACGCCCCGTGTCTGGAT
GCACAGATGCCATGGCCTGTGCTGGGCCAGTGGCTGGGGGTGCTAGACACCCGGCACCATTCTCCCTTCT
CTCTTTTCTTCTCAGGATTTAAAATTTAATTATATCAGTAAAGAGATTAATTTTAACGTAACTCTTTCTA
TGCCCGTGTAAAGTATGTGAATCGCAAGGCCTGTGCTGCATGCGACAGCGTCCGGGGTGGTGGACAGGGC
CCCCGGCCACGCTCCCTCTCCTGTAGCCACTGGCATAGCCCTCCTGAGCACCCGCTGACATTTCCGTTGT
ACATGTTCCTGTTTATGCATTCACAAGGTGACTGGGATGTAGAGAGGCGTTAGTGGGCAGGTGGCCACAG
CAGGACTGAGGACAGGCCCCCATTATCCTAGGGGTGCGCTCACCTGCAGCCCCTCCTCCTCGGGCACAGA
CGACTGTCGTTCTCCACCCACCAGTCAGGGACAGCAGCCTCCCTGTCACTCAGCTGAGAAGGCCAGCCCT
CCCTGGCTGTGAGCAGCCTCCACTGTGTCCAGAGACATGGGCCTCCCACTCCTGTTCCTTGCTAGCCCTG
GGGTGGCGTCTGCCTAGGAGCTGGCTGGCAGGTGTTGGGACCTGCTGCTCCATGGATGCATGCCCTAAGA
GTGTCACTGAGCTGTGTTTTGTCTGAGCCTCTCTCGGTCAACAGCAAAGCTTGGTGTCTTGGCACTGTTA
GTGACAGAGCCCAGCATCCCTTCTGCCCCCGTTCCAGCTGACATCTTGCACGGTGACCCCTTTTAGTCAG
GAGAGTGCAGATCTGTGCTCATCGGAGACTGCCCCACGGCCCTGTCAGAGCCGCCACTCCTATCCCCAGG
CCAGGTCCCTGGACCAGCCTCCTGTTTGCAGGCCCAGAGGAGCCAAGTCATTAAAATGGAAGTGGATTCT
GGATGGCCGGGCTGCTGCTGATGTAGGAGCTGGATTTGGGAGCTCTGCTTGCCGACTGGCTGTGAGACGA
GGCAGGGGCTCTGCTTCCTCAGCCCTAGAGGCGAGCCAGGCAAGGTTGGCGACTGTCATGTGGCTTGGTT
TGGTCATGCCCGTCGATGTTTTGGGTATTGAATGTGGTAAGTGGAGGAAATGTTGGAACTCTGTGCAGGT
GCTGCCTTGAGACCCCCAAGCTTCCACCTGTCCCTCTCCTATGTGGCAGCTGGGGAGCAGCTGAGATGTG
GACTTGTATGCTGCCCACATACGTGAGGGGGAGCTGAAAGGGAGCCCCTCCTCTGAGCAGCCTCTGCCAG
GCCTGTATGAGGCTTTTCCCACCAGCTCCCAACAGAGGCCTCCCCCAGCCAGGACCACCTCGTCCTCGTG
GCGGGGCAGCAGGAGCGGTAGAAAGGGGTCCGATGTTTGAGGAGGCCCTTAAGGGAAGCTACTGAATTAT
AACACGTAAGAAAATCACCATTCCGTATTGGTTGGGGGCTCCTGTTTCTCATCCTAGCTTTTTCCTGGAA
AGCCCGCTAGAAGGTTTGGGAACGAGGGGAAAGTTCTCAGAACTGTTGGCTGCTCCCCACCCGCCTCCCG
CCTCCCCCGCAGGTTATGTCAGCAGCTCTGAGACAGCAGTATCACAGGCCAGATGTTGTTCCTGGCTAGA
TGTTTACATTTGTAAGAAATAACACTGTGAATGTAAAACAGAGCCATTCCCTTGGAATGCATATCGCTGG
GCTCAACATAGAGTTTGTCTTCCTCTTGTTTACGACGTGATCTAAACCAGTCCTTAGCAAGGGGCTCAGA
ACACCCCGCTCTGGCAGTAGGTGTCCCCCACCCCCAAAGACCTGCCTGTGTGCTCCGGAGATGAATATGA
GCTCATTAGTAAAAATGACTTCACCCACGCATATACATAAAGTATCCATGCATGTGCATATAGACACATC
TATAATTTTACACACACACCTCTCAAGACGGAGATGCATGGCCTCTAAGAGTGCCCGTGTCGGTTCTTCC
TGGAAGTTGACTTTCCTTAGACCCGCCAGGTCAAGTTAGCCGCGTGACGGACATCCAGGCGTGGGACGTG
GTCAGGGCAGGGCTCATTCATTGCCCACTAGGATCCCACTGGCGAAGATGGTCTCCATATCAGCTCTCTG
CAGAAGGGAGGAAGACTTTATCATGTTCCTAAAAATCTGTGGCAAGCACCCATCGTATTATCCAAATTTT
GTTGCAAATGTGATTAATTTGGTTGTCAAGTTTTGGGGGTGGGCTGTGGGGAGATTGCTTTTGTTTTCCT
GCTGGTAATATCGGGAAAGATTTTAATGAAACCAGGGTAGAATTGTTTGGCAATGCACTGAAGCGTGTTT
CTTTCCCAAAATGTGCCTCCCTTCCGCTGCGGGCCCAGCTGAGTCTATGTAGGTGATGTTTCCAGCTGCC
AAGTGCTCTTTGTTACTGTCCACCCTCATTTCTGCCAGCGCATGTGTCCTTTCAAGGGGAAAATGTGAAG
CTGAACCCCCTCCAGACACCCAGAATGTAGCATCTGAGAAGGCCCTGTGCCCTAAAGGACACCCCTCGCC
CCCATCTTCATGGAGGGGGTCATTTCAGAGCCCTCGGAGCCAATGAACAGCTCCTCCTCTTGGAGCTGAG
ATGAGCCCCACGTGGAGCTCGGGACGGATAGTAGACAGCAATAACTCGGTGTGTGGCCGCCTGGCAGGTG
GAACTTCCTCCCGTTGCGGGGTGGAGTGAGGTTAGTTCTGTGTGTCTGGTGGGTGGAGTCAGGCTTCTCT
TGCTACCTGTGAGCATCCTTCCCAGCAGACATCCTCATCGGGCTTTGTCCCTCCCCCGCTTCCTCCCTCT
GCGGGGAGGACCCGGGACCACAGCTGCTGGCCAGGGTAGACTTGGAGCTGTCCTCCAGAGGGGTCACGTG
TAGGAGTGAGAAGAAGGAAGATCTTGAGAGCTGCTGAGGGACCTTGGAGAGCTCAGGATGGCTCAGACGA
GGACACTCGCTTGCCGGGCCTGGGCCTCCTGGGAAGGAGGGAGCTGCTCAGAATGCCGCATGACAACTGA
AGGCAACCTGGAAGGTTCAGGGGCCGCTCTTCCCCCATGTGCCTGTCACGCTCTGGTGCAGTCAAAGGAA
CGCCTTCCCCTCAGTTGTTTCTAAGAGCAGAGTCTCCCGCTGCAATCTGGGTGGTAACTGCCAGCCTTGG
AGGATCGTGGCCAACGTGGACCTGCCTACGGAGGGTGGGCTCTGACCCAAGTGGGGCCTCCTTGTCCAGG
TCTCACTGCTTTGCACCGTGGTCAGAGGGACTGTCAGCTGAGCTTGAGCTCCCCTGGAGCCAGCAGGGCT
GTGATGGGCGAGTCCCGGAGCCCCACCCAGACCTGAATGCTTCTGAGAGCAAAGGGAAGGACTGACGAGA
GATGTATATTTAATTTTTTAACTGCTGCAAACATTGTACATCCAAATTAAAGGAAAAAAATGGAAACCAT
CAAAAAAAAAAAAAAAAAA

Referências.

Hermankova M, Ray SC, Ruff C, et al. Hiv- 1 drug resistance profiles in children and adults with viral load of <50 copies/ml receiving combination therapy. JAMA. 2001;286:196–207. [PubMed]