Bioinformática

COMO UTILIZAR O GENEBANK

NCBI -National Center for Biotechnology Information

Costa de Oliveira, Antonio

Maia, Luciano Carlos da

Centro de Genômica e Fitomelhoramento

FAEM - Universidade Federal Pelotas
 BLAST - Basic Local Alignment Search Tool
Altschul et al, 1990

Exercicio 1 BLASTn

Nucleotide query x Nucleoide DataBase

Search 5’ and/or reverse complement sequence
NCBI -National Center for Biotechnology Information

Exercicio 1: Como utilizar o BLAST no NCBI (on-line).

BLASTn (nucleotideos x nucleotideos)

A. Dada uma sequencia de nucleotideos, verificar se existem sequencias homologas

e quais as informaçoes referenciadas para suas “homologias”

A1. Interpretando o E-value

A2. Interpretaçao detalhada dos alinhamentos com melhor E-value

A3. Existem informaçoes genomicas para os homologos?

BACs?
Genoma completo?
Qual a estrutura deste gene?
promoter
introns
exons (UTR, CDS)

A4. Existem informações de homologias em bancos de dados de regioes

expressas?
A5. Exitem informações sobre a validação experimental destes homologos?
NCBI -National Center for Biotechnology Information

Exercicio 1: Como utilizar o BLAST no NCBI (on-line).

BLASTn (nucleotideos x nucleotideos)

ATGGGGGTGACGAAGGCGGAGGCGGTTGCCGCGACGGGGAAGGTGGTGGACGACATCGAGGCGC
TCGCCGATCTCAGAAAGGAGCCAGCATGGAAAAAGTTTTTGTCCCACATTGGACCAGGCTTTATGGT
CTGTTTGGCTTACCTCGATCCTGGAAATATGGAGACTGATCTACAAGCTGGTGCCAATCACAAATATG
AGCTTCTTTGGGTGATTTTGATTGGCCTGATCTTTGCACTGATTATACAATCATTATCGGCTAATCTTG
GAGTGGTGACAGGGCGCCATCTTGCTGAGCTGTGCAAGACTGAATATCCAGTATGGGTGAAAACCT
GCTTATGGCTGCTAGCAGAACTAGCTGTGATTGCTTCCGATATTCCAGAAGTTATAGGTACAGGATTT
GCTTTCAACCTTTTGTTCCACATACCAGTGTGGACCGGGGTTCTCATTGCTGGCTCTAGCACGCTTC
TTCTTCTTGGACTGCAAAGATATGGGGTAAGGAAACTGGAGGTTGTGGTCGCGCTATTGGTGTTTGT
CATGGCAGGGTGCTTCTTCGTGGAGATGAGCATAGTCAAGCCTCCGGTTAATGAGGTCCTTCAAGG
ATTATTCATCCCCAGGCTCAGTGGGCCCGGTGCCACAGGAGACTCCATTGCCCTTCTTGGGGCTCTT
GTAATGCCGCACAATCTATTCTTGCACTCTGCCCTGGTGCTGTCGAGGAATACACCCGCATCAGCAA
AAGGAATGAAGGATGCCTGTAGGTTCTTCCTCTTTGAGAGTGGGATAGCTCTTTTCGTGGCTCTGCT
TGTCAACATTGCAATCATCTCTGTCTCCGGCACTGTATGTAATGCAACCAACCTTTCACCAGAAGATG
CTGTAAAATGCAGCGACCTTACATTGGACTCCTCATCCTTCCTCCTCAGGAATGTGCTAGGAAAGTCT
AGTGCGACAGTGTACGGTGTAGCACTCTTGGCTTCTGGACAGAGCTCGACCATTACTGGCACTTATG
CTGGGCAATATGTTATGCAGGGGTTCCTAGACATCAAAATGAAACAGTGGCTTCGGAACCTGATGAC
ACGCAGCATTGCCATTGTGCCTAGCTTAATCGTCTCCATCATTGGAGGGTCTAGTGGCGCCGGTCGT
CTCATCGTCATTGCATCGATGATTCTATCCTTTGAGCTACCATTTGCTCTGATCCCTCTTCTCAAGTTC
AGCAGCAGCAGCAACAAGATGGGTGAAAACAAGAACTCCATCTATATTGTTGGATTCTCCTGGGTGC
TGGGGTTCGTCATCATCGGGATAAACATCTACTTCCTCAGCACAAAGCTGGTCGGCTGGATCCTCCA
CAACGCGCTCCCGACATTCGCCAACGTGCTCATCGGCATCGTGCTGTTCCCGCTCATGCTGCTCTA
CGTCGTCGCCGTGATCTACCTGACGTTCAGGAAGGACACCGTCAAGTTCGTGTCTCGTCGTGAGCT
GCAGGCCGGCGACGACACCGAGAAAGCACAGGTAGCCACCTGCGTCGCCGACGAGGACAGCAAA
G AGCCACCCGTGTAG
ATGGGGGTGACGAAGGCGGAGGCGGTTGCCGCGACGGGGAAGGTGGTGGACGACATCGAGGCGC
TCGCCGATCTCAGAAAGGAGCCAGCATGGAAAAAGTTTTTGTCCCACATTGGACCAGGCTTTATGGT
 BLAST - Basic Local Alignment Search Tool
Altschul et al, 1990
Exercicio 2

BLASTx

Nucleotide query x Protein DataBase (a.á)

Search nucleotide sequence translated in all 6 reading frame
NCBI -National Center for Biotechnology Information

Exercicio 2: Como utilizar o BLAST no NCBI (on-line).

BLASTx (nucleotideos x amino ácidos)

B. Dada uma sequencia de nucleotideos, verificar se existem sequencias proteicas

(a.á) homologas e quais as informaçoes referenciadas para suas “homologias”

B1. Interpretando o E-value

B2. Interpretaçao detalhada dos alinhamentos com melhor E-value

B3. Existem informaçoes genomicas para os homologos?

BACs?
Genoma completo?
Qual a estrutura deste gene?
promoter
introns
exons (UTR, CDS)

B4. Existem informações de homologias em bancos de dados de regioes

expressas?
B5. Exitem informações sobre a validação experimental destes homologos?
B6. Existem informação estrutural desta proteina? Cristalografia?
NCBI -National Center for Biotechnology Information

Exercicio 2: Como utilizar o BLAST no NCBI (on-line).

BLASTx (nucleotideos x amino ácidos)

ATGGCGTCGAAGGAGGAGGTGGCCGTGGAGACGGTGGAGGGCGGAG
CGGCGGCGGCGAAGGCGCCGTACTGGACCCGCCGCCGGCGCCGCTG
CTGGACACGAGCGAGCTGGGTAAGTGGTCGCTGTACCGTGCCCTCAT
CGCGGAGTTCATGGCGACGCTCATCTTCCTCTACGTGAGCATCGCCAC
CGTGATCGGGTACAAGAACCAGAGGGCCACCGTCGACGCGTGCACCG
GCGTCGGCTACCTCGGCGTGGCGTGGTCGTTCGGTGCCACCATAT
TCGTCCTCGTCTACTGCACCGGCGGCGTCTCCGGCGGGCACATCAACC
CGGCGGTGACGCTGGGCCTCTTCTTCGGGCGGAAGCTGTCGCTCGTC
CGCACCGTGCTGTACGTCGTGGCGCAGTGCCTCGGCGCCATCGCC
GGCGCCGGCATCGTCGGCACGTTCATCCTCGTCTACACCGTCTTCTCC
GCCACCGACCCCAAGCGCACCGCCCGCGACTCCTTCATCCCCGTACTC
GTGCCGCTGCCGATCGGGTTCGCGGTGTTCGTCGTGCACCTGGC
GACGATTCCGATCACCGGCACGGGCATCAACCCGGCGAGGAGCCTCG
GCGCCGCCGTGCTGTACAACCAGCACGCAGCTTGGAAAGACCACTGGA
TCTTCTGGGTGGGGCCGGTGATCGGGGCGTTCTTGGCGGCGGCGT
ACCACAAGCTGGTGCTGCGCGGCGAGGCCGCCAAGGCGCTCAGCTCG
TTCAGGAGCACCAGCGTGACGGCGTGA
 BLAST - Basic Local Alignment Search Tool
Altschul et al, 1990
Exercicio 3
BLASTp

Protein (a.á) query x Protein (a.á) DataBase

NCBI -National Center for Biotechnology Information

Exercicio 3: Como utilizar o BLAST no NCBI (on-line).

BLASTp (aminóácidox aminoácidos)

LGVPSVKHMFPFCDSSSPMDLPLYQQLQLSPSSPKTDQSSSFYC
YPCSPPFAAADASFPLSYQIGSAAAADATPPQAVINSPDLPVQALMDHAPA
TATELGACASGAEGSGASLDRAAAAARKDRHSKICTAGGMRDRRMRLSLD
VARKFFALQDMLGFDKASKTVQWLLNTSKSAIQEIMADDASSECVEDGSSS
LSVDGKHNPAEQLGGGGDQKPKGNCRGEGKKPAKASKAAATPKPPRKSA
NNAHQVPDKETRAKARERARERTKEKHRMRWVKLASAIDVEAAAASGPSD
RPSSNNLSHHSSLSMNMPCAAAELEERERCSSPLSNRSAGRMQEITGASD
VVLGFGNGGGGYGDGGGNYYCQEQWELGGVVFQQNSRFY
 BLAST - Basic Local Alignment Search Tool
Altschul et al, 1990

Exercicio 4 TBLASTn

Protein (a.á) query x Nucleotide DataBase

Translated in all Reading Frame
NCBI -National Center for Biotechnology Information

Exercicio 4: Como utilizar o BLAST no NCBI (on-line).

TBLASTn (aminóácidox nucleotideos)

LGVPSVKHMFPFCDSSSPMDLPLYQQLQLSPSSPKTDQSSSFYC
YPCSPPFAAADASFPLSYQIGSAAAADATPPQAVINSPDLPVQALMDHAPA
TATELGACASGAEGSGASLDRAAAAARKDRHSKICTAGGMRDRRMRLSLD
VARKFFALQDMLGFDKASKTVQWLLNTSKSAIQEIMADDASSECVEDGSSS
LSVDGKHNPAEQLGGGGDQKPKGNCRGEGKKPAKASKAAATPKPPRKSA
NNAHQVPDKETRAKARERARERTKEKHRMRWVKLASAIDVEAAAASGPSD
RPSSNNLSHHSSLSMNMPCAAAELEERERCSSPLSNRSAGRMQEITGASD
VVLGFGNGGGGYGDGGGNYYCQEQWELGGVVFQQNSRFY
 BLAST - Basic Local Alignment Search Tool
Altschul et al, 1990
Exercicio 5

TBLASTx

Nucleotide translated query x Nucleotide DataBase (Nucleotide translated query)

Translated in 6 all Reading Frame x Translated in 6 all Reading Frame
NCBI -National Center for Biotechnology Information

Exercicio 5: Como utilizar o BLAST no NCBI (on-line).

TBLASTn (aminóácidox nucleotideos)

ATGGAAACTGGCAACGTTGTGAATGCACAACCTGAATTATCAGGCATAATTGATGGATCCAAGAGCTATA
TGTACGAGCAGTTATGGAAACTCTGTGCGGGACCTTTGTGTGATATTCCAAAACTTGGAGAAAATGTTTA
TTACTTTCCTCAAGGCAATATAGAGCTTGTTGATGCATCTACAAGAGAAGAGTTGAATGAGTTACAACCA
ATTTGTGATCTTCCTTCTAAGCTTCAATGCCGTGTTATTGCAATTCATCTTAAGGTGGAAAACAATAGTG
ATGAGACTTATGCTGAGATCACTTTGATGCCAGATACAACTCAAGTTGTAATCCCTACTCAGAGTGAAAA
TCAATTTAGGCCATTAGTTAATTCCTTTACGAAGGTTTTAACGGCCTCTGATACAAGCGCTTATGGTGGA
TTCTTTGTTCCGAAAAAACATGCCATTGAATGTCTCCCTCCACTGCCTCTACCGGCACAAGAACTTCTTG
CTAAAGATCTCCATGGTAATCAATGGAGATTCAGACACAGCTATAGAGGTACACCGCAAAGACATTCGTT
AACAACTGGTTGGAATGAATTCACAACATCGAAAAAGTTGGTTAAAGGAGATGTTATTGTATTCGTTAGG
GGAGAGACTGGAGAGTTACGAGTTGGTATCAGACGAGCAAGACATCAACAAGGGAATATACCTTCATCAA
TAGTATCAATAGATTGTATGAGACATGGGGTAATTGCTTCCGCAAAGCATGCTCTTGATAACCAATGTAT
ATTCATTGTGGTTTACAAGCCAAGTATCAGGTCAAGTCAATTTATTGTCAGTTACGACAAATTCTTAGAC
GCTATGAACAATAAGTTCATTGTCGGTTCAAGATTTACAATGCGGTTTGAGGGTGATGATTTCTCTGAAA
GAAGATATTTTGGGACAATTATTGGAGTTAATGATTTCTCTCCTCATTGGAAGTGTTCAGAGTGGCGCAG
CTTAGAAGTTCAGTGGGATGAATTTGCATCATTTTCGAGACCTAATAAAGTGTCACCCTGGGAGATCGAA
CATTTAATGTCTGCGTTAAATGTTCCGCGATCATCTTTGCTCAAGAACAAGCGTTTACGAGAAGTTAATG
AATTCGGACAAGAAATTGGACAACTAAGCGTCGCCTCCCCGATGAATACTTCTCTTCGTTATCGTGATAC
AACTGAAGATGCTATGAATCCTTCTAGATTGCTAATGAGCTACCCTGTCCAACCAATGCCCAAACTAAAT
TACAATAACCAAATGGTTACACAAATAGAAGAAAATATAACAACCAAGGCAGTCACTAATTTTAGGCTGT
TTGGAGTCTCTCTAGCCATTCCTCTAGTGATCAAAGATCCCATTGAAGAAATTGGCTCGGATATTTCGAA
ACTTACTGAAGGAAAAAAGTTTGGTCAAAGCCAAACTTTGAGATCACCAATAGAGATCCAAAGCAAGCAG
TTTGGTTCTACTAGAACTTGTACCAAAGTTCAAATGCAAGGTGTAACAATAGGAAGAGCTGTGGATTTAA
GTGTTCTTAATGGATATGATCAGCTAATACTAGAATTGGAGAAGCTCTTTGATCTCAAAGGCCAACTACA
AACTCGCAACCAATGGAAAATAGCTTTCACAGATAGTGACGGGTATGAGATGCTTGTTGGAGACGATCCA
TGGCCTGAATTCTGCAAAATGGTGAAGAAGATATTGATATATTCAAAAGAGGAGGTCAAAAACCTAAAGT
CCAGTAAGAGTCTCTCAAGTTGA
??????????????
 TRABALHO PARA PRÓXIMA AULA

1) BUSCAR HOMOLOGIAS (BLAST)

JULGAR QUAL SEQUENCIAS SAO AS MAIS PARECIDAS

2) DESCRICAO DA FUNÇÃO DO GENE/PROTEÍNA

3) EXISTE SEQUENCIAS CONFIRMANDO A EXPRESSÃO DE HOMOLOGOS

DESTAS SEQUENCIAS??
UNIGENE

4) EXISTE INFORMAÇÃO DA VALIDAÇÃO EXPERIMENTAL DOS HOMOLOGOS

DESTA SEQUENCIA? EM QUE ESPECIE? SAO RESPONSIVOS A QUAIS
FATORES?
GEOPROFILE

5) EXPLIQUE COMO QUANTO E PARA QUE ESSE GENE PODE SER UTILIZADO
COMO MARCADOR MOLECULAR!!!
 TRABALHO PARA PRÓXIMA AULA

RESULTADO DO TRABALHO:

1)OS DADOS SERAO APRESENTADOS NA PROXIMA AULA POR CADA DUPLA

2) APRESENTAR EM 15 MIN NO POWERPOINT

 Bioinformática

COMO UTILIZAR O GENEBANK

NCBI -National Center for Biotechnology Information

Costa de Oliveira, Antonio

Maia, Luciano Carlos da

Centro de Genômica e Fitomelhoramento

FAEM - Universidade Federal Pelotas
 BLAST - Basic Local Alignment Search Tool
Altschul et al, 1990

Exercicio 1 BLASTn

Nucleotide query x Nucleoide DataBase

Search 5’ and/or reverse complement sequence
NCBI -National Center for Biotechnology Information

Exercicio 1: Como utilizar o BLAST no NCBI (on-line).

BLASTn (nucleotideos x nucleotideos)

A. Dada uma sequencia de nucleotideos, verificar se existem sequencias homologas

e quais as informaçoes referenciadas para suas “homologias”

A1. Interpretando o E-value

A2. Interpretaçao detalhada dos alinhamentos com melhor E-value

A3. Existem informaçoes genomicas para os homologos?

BACs?
Genoma completo?
Qual a estrutura deste gene?
promoter
introns
exons (UTR, CDS)

A4. Existem informações de homologias em bancos de dados de regioes

expressas?
A5. Exitem informações sobre a validação experimental destes homologos?
NCBI -National Center for Biotechnology Information

Exercicio 1: Como utilizar o BLAST no NCBI (on-line).

BLASTn (nucleotideos x nucleotideos)

ATGGGGGTGACGAAGGCGGAGGCGGTTGCCGCGACGGGGAAGGTGGTGGACGACATCGAGGCGC
TCGCCGATCTCAGAAAGGAGCCAGCATGGAAAAAGTTTTTGTCCCACATTGGACCAGGCTTTATGGT
CTGTTTGGCTTACCTCGATCCTGGAAATATGGAGACTGATCTACAAGCTGGTGCCAATCACAAATATG
AGCTTCTTTGGGTGATTTTGATTGGCCTGATCTTTGCACTGATTATACAATCATTATCGGCTAATCTTG
GAGTGGTGACAGGGCGCCATCTTGCTGAGCTGTGCAAGACTGAATATCCAGTATGGGTGAAAACCT
GCTTATGGCTGCTAGCAGAACTAGCTGTGATTGCTTCCGATATTCCAGAAGTTATAGGTACAGGATTT
GCTTTCAACCTTTTGTTCCACATACCAGTGTGGACCGGGGTTCTCATTGCTGGCTCTAGCACGCTTC
TTCTTCTTGGACTGCAAAGATATGGGGTAAGGAAACTGGAGGTTGTGGTCGCGCTATTGGTGTTTGT
CATGGCAGGGTGCTTCTTCGTGGAGATGAGCATAGTCAAGCCTCCGGTTAATGAGGTCCTTCAAGG
ATTATTCATCCCCAGGCTCAGTGGGCCCGGTGCCACAGGAGACTCCATTGCCCTTCTTGGGGCTCTT
GTAATGCCGCACAATCTATTCTTGCACTCTGCCCTGGTGCTGTCGAGGAATACACCCGCATCAGCAA
AAGGAATGAAGGATGCCTGTAGGTTCTTCCTCTTTGAGAGTGGGATAGCTCTTTTCGTGGCTCTGCT
TGTCAACATTGCAATCATCTCTGTCTCCGGCACTGTATGTAATGCAACCAACCTTTCACCAGAAGATG
CTGTAAAATGCAGCGACCTTACATTGGACTCCTCATCCTTCCTCCTCAGGAATGTGCTAGGAAAGTCT
AGTGCGACAGTGTACGGTGTAGCACTCTTGGCTTCTGGACAGAGCTCGACCATTACTGGCACTTATG
CTGGGCAATATGTTATGCAGGGGTTCCTAGACATCAAAATGAAACAGTGGCTTCGGAACCTGATGAC
ACGCAGCATTGCCATTGTGCCTAGCTTAATCGTCTCCATCATTGGAGGGTCTAGTGGCGCCGGTCGT
CTCATCGTCATTGCATCGATGATTCTATCCTTTGAGCTACCATTTGCTCTGATCCCTCTTCTCAAGTTC
AGCAGCAGCAGCAACAAGATGGGTGAAAACAAGAACTCCATCTATATTGTTGGATTCTCCTGGGTGC
TGGGGTTCGTCATCATCGGGATAAACATCTACTTCCTCAGCACAAAGCTGGTCGGCTGGATCCTCCA
CAACGCGCTCCCGACATTCGCCAACGTGCTCATCGGCATCGTGCTGTTCCCGCTCATGCTGCTCTA
CGTCGTCGCCGTGATCTACCTGACGTTCAGGAAGGACACCGTCAAGTTCGTGTCTCGTCGTGAGCT
GCAGGCCGGCGACGACACCGAGAAAGCACAGGTAGCCACCTGCGTCGCCGACGAGGACAGCAAA
G AGCCACCCGTGTAG
 BLAST - Basic Local Alignment Search Tool
Altschul et al, 1990
Exercicio 2

BLASTx

Nucleotide query x Protein DataBase (a.á)

Search nucleotide sequence translated in all 6 reading frame
NCBI -National Center for Biotechnology Information

Exercicio 2: Como utilizar o BLAST no NCBI (on-line).

BLASTx (nucleotideos x amino ácidos)

B. Dada uma sequencia de nucleotideos, verificar se existem sequencias proteicas

(a.á) homologas e quais as informaçoes referenciadas para suas “homologias”

B1. Interpretando o E-value

B2. Interpretaçao detalhada dos alinhamentos com melhor E-value

B3. Existem informaçoes genomicas para os homologos?

BACs?
Genoma completo?
Qual a estrutura deste gene?
promoter
introns
exons (UTR, CDS)

B4. Existem informações de homologias em bancos de dados de regioes

expressas?
B5. Exitem informações sobre a validação experimental destes homologos?
B6. Existem informação estrutural desta proteina? Cristalografia?
NCBI -National Center for Biotechnology Information

Exercicio 2: Como utilizar o BLAST no NCBI (on-line).

BLASTx (nucleotideos x amino ácidos)

ATGGCGTCGAAGGAGGAGGTGGCCGTGGAGACGGTGGAGGGCGGAG
CGGCGGCGGCGAAGGCGCCGTACTGGACCCGCCGCCGGCGCCGCTG
CTGGACACGAGCGAGCTGGGTAAGTGGTCGCTGTACCGTGCCCTCAT
CGCGGAGTTCATGGCGACGCTCATCTTCCTCTACGTGAGCATCGCCAC
CGTGATCGGGTACAAGAACCAGAGGGCCACCGTCGACGCGTGCACCG
GCGTCGGCTACCTCGGCGTGGCGTGGTCGTTCGGTGCCACCATAT
TCGTCCTCGTCTACTGCACCGGCGGCGTCTCCGGCGGGCACATCAACC
CGGCGGTGACGCTGGGCCTCTTCTTCGGGCGGAAGCTGTCGCTCGTC
CGCACCGTGCTGTACGTCGTGGCGCAGTGCCTCGGCGCCATCGCC
GGCGCCGGCATCGTCGGCACGTTCATCCTCGTCTACACCGTCTTCTCC
GCCACCGACCCCAAGCGCACCGCCCGCGACTCCTTCATCCCCGTACTC
GTGCCGCTGCCGATCGGGTTCGCGGTGTTCGTCGTGCACCTGGC
GACGATTCCGATCACCGGCACGGGCATCAACCCGGCGAGGAGCCTCG
GCGCCGCCGTGCTGTACAACCAGCACGCAGCTTGGAAAGACCACTGGA
TCTTCTGGGTGGGGCCGGTGATCGGGGCGTTCTTGGCGGCGGCGT
ACCACAAGCTGGTGCTGCGCGGCGAGGCCGCCAAGGCGCTCAGCTCG
TTCAGGAGCACCAGCGTGACGGCGTGA
 BLAST - Basic Local Alignment Search Tool
Altschul et al, 1990
Exercicio 3
BLASTp

Protein (a.á) query x Protein (a.á) DataBase

NCBI -National Center for Biotechnology Information

Exercicio 3: Como utilizar o BLAST no NCBI (on-line).

BLASTp (aminóácidox aminoácidos)

LGVPSVKHMFPFCDSSSPMDLPLYQQLQLSPSSPKTDQSSSFYC
YPCSPPFAAADASFPLSYQIGSAAAADATPPQAVINSPDLPVQALMDHAPA
TATELGACASGAEGSGASLDRAAAAARKDRHSKICTAGGMRDRRMRLSLD
VARKFFALQDMLGFDKASKTVQWLLNTSKSAIQEIMADDASSECVEDGSSS
LSVDGKHNPAEQLGGGGDQKPKGNCRGEGKKPAKASKAAATPKPPRKSA
NNAHQVPDKETRAKARERARERTKEKHRMRWVKLASAIDVEAAAASGPSD
RPSSNNLSHHSSLSMNMPCAAAELEERERCSSPLSNRSAGRMQEITGASD
VVLGFGNGGGGYGDGGGNYYCQEQWELGGVVFQQNSRFY
 BLAST - Basic Local Alignment Search Tool
Altschul et al, 1990

Exercicio 4 TBLASTn

Protein (a.á) query x Nucleotide DataBase

Translated in all Reading Frame
NCBI -National Center for Biotechnology Information

Exercicio 4: Como utilizar o BLAST no NCBI (on-line).

TBLASTn (aminóácidox nucleotideos)

LGVPSVKHMFPFCDSSSPMDLPLYQQLQLSPSSPKTDQSSSFYC
YPCSPPFAAADASFPLSYQIGSAAAADATPPQAVINSPDLPVQALMDHAPA
TATELGACASGAEGSGASLDRAAAAARKDRHSKICTAGGMRDRRMRLSLD
VARKFFALQDMLGFDKASKTVQWLLNTSKSAIQEIMADDASSECVEDGSSS
LSVDGKHNPAEQLGGGGDQKPKGNCRGEGKKPAKASKAAATPKPPRKSA
NNAHQVPDKETRAKARERARERTKEKHRMRWVKLASAIDVEAAAASGPSD
RPSSNNLSHHSSLSMNMPCAAAELEERERCSSPLSNRSAGRMQEITGASD
VVLGFGNGGGGYGDGGGNYYCQEQWELGGVVFQQNSRFY
 BLAST - Basic Local Alignment Search Tool
Altschul et al, 1990
Exercicio 5

TBLASTx

Nucleotide translated query x Nucleotide DataBase (Nucleotide translated query)

Translated in 6 all Reading Frame x Translated in 6 all Reading Frame
NCBI -National Center for Biotechnology Information

Exercicio 5: Como utilizar o BLAST no NCBI (on-line).

TBLASTn (aminóácidox nucleotideos)

ATGGAAACTGGCAACGTTGTGAATGCACAACCTGAATTATCAGGCATAATTGATGGATCCAAGAGCTATA
TGTACGAGCAGTTATGGAAACTCTGTGCGGGACCTTTGTGTGATATTCCAAAACTTGGAGAAAATGTTTA
TTACTTTCCTCAAGGCAATATAGAGCTTGTTGATGCATCTACAAGAGAAGAGTTGAATGAGTTACAACCA
ATTTGTGATCTTCCTTCTAAGCTTCAATGCCGTGTTATTGCAATTCATCTTAAGGTGGAAAACAATAGTG
ATGAGACTTATGCTGAGATCACTTTGATGCCAGATACAACTCAAGTTGTAATCCCTACTCAGAGTGAAAA
TCAATTTAGGCCATTAGTTAATTCCTTTACGAAGGTTTTAACGGCCTCTGATACAAGCGCTTATGGTGGA
TTCTTTGTTCCGAAAAAACATGCCATTGAATGTCTCCCTCCACTGCCTCTACCGGCACAAGAACTTCTTG
CTAAAGATCTCCATGGTAATCAATGGAGATTCAGACACAGCTATAGAGGTACACCGCAAAGACATTCGTT
AACAACTGGTTGGAATGAATTCACAACATCGAAAAAGTTGGTTAAAGGAGATGTTATTGTATTCGTTAGG
GGAGAGACTGGAGAGTTACGAGTTGGTATCAGACGAGCAAGACATCAACAAGGGAATATACCTTCATCAA
TAGTATCAATAGATTGTATGAGACATGGGGTAATTGCTTCCGCAAAGCATGCTCTTGATAACCAATGTAT
ATTCATTGTGGTTTACAAGCCAAGTATCAGGTCAAGTCAATTTATTGTCAGTTACGACAAATTCTTAGAC
GCTATGAACAATAAGTTCATTGTCGGTTCAAGATTTACAATGCGGTTTGAGGGTGATGATTTCTCTGAAA
GAAGATATTTTGGGACAATTATTGGAGTTAATGATTTCTCTCCTCATTGGAAGTGTTCAGAGTGGCGCAG
CTTAGAAGTTCAGTGGGATGAATTTGCATCATTTTCGAGACCTAATAAAGTGTCACCCTGGGAGATCGAA
CATTTAATGTCTGCGTTAAATGTTCCGCGATCATCTTTGCTCAAGAACAAGCGTTTACGAGAAGTTAATG
AATTCGGACAAGAAATTGGACAACTAAGCGTCGCCTCCCCGATGAATACTTCTCTTCGTTATCGTGATAC
AACTGAAGATGCTATGAATCCTTCTAGATTGCTAATGAGCTACCCTGTCCAACCAATGCCCAAACTAAAT
TACAATAACCAAATGGTTACACAAATAGAAGAAAATATAACAACCAAGGCAGTCACTAATTTTAGGCTGT
TTGGAGTCTCTCTAGCCATTCCTCTAGTGATCAAAGATCCCATTGAAGAAATTGGCTCGGATATTTCGAA
ACTTACTGAAGGAAAAAAGTTTGGTCAAAGCCAAACTTTGAGATCACCAATAGAGATCCAAAGCAAGCAG
TTTGGTTCTACTAGAACTTGTACCAAAGTTCAAATGCAAGGTGTAACAATAGGAAGAGCTGTGGATTTAA
GTGTTCTTAATGGATATGATCAGCTAATACTAGAATTGGAGAAGCTCTTTGATCTCAAAGGCCAACTACA
AACTCGCAACCAATGGAAAATAGCTTTCACAGATAGTGACGGGTATGAGATGCTTGTTGGAGACGATCCA
TGGCCTGAATTCTGCAAAATGGTGAAGAAGATATTGATATATTCAAAAGAGGAGGTCAAAAACCTAAAGT
CCAGTAAGAGTCTCTCAAGTTGA
??????????????
 TRABALHO PARA PRÓXIMA AULA

1) BUSCAR HOMOLOGIAS (BLAST)

JULGAR QUAIS SEQUENCIAS SAO AS MAIS PARECIDAS!

2) DESCRICAO DA FUNÇÃO DO GENE/PROTEÍNA

3) EXISTEM SEQUENCIAS CONFIRMANDO A EXPRESSÃO PARA OS

HOMOLÓGOS DESTA SEQUENCIA??
VER SE EXISTEM HOMOLÓGOS NO UNIGENE (cDNA)

4) EXISTEM INFORMAÇÕES DA VALIDAÇÃO EXPERIMENTAL DOS HOMOLOGOS

DESTA SEQUENCIA? EM QUE ESPECIE? SÃO RESPONSIVOS A QUAIS
FATORES?
GEOPROFILE

5) EXPLIQUE COMO, QUANTO E PARA QUE ESSE GENE (OU SEQUENCIA

GENOMICA) PODERA SER UTILIZADO/A COMO MARCADOR MOLECULAR!!!
 TRABALHO PARA PRÓXIMA AULA

RESULTADO DO TRABALHO:

1)OS DADOS SERÃO APRESENTADOS NA PRÓXIMA AULA POR CADA DUPLA

2) APRESENTAR EM 15 MINUTOS NO POWERPOINT