Ljubica Tasic

ljubica@iqm.unicamp.br I-249 e I-250 F: 11106 e 13042

Clonagem Expresso Purificao Anlises estruturais RMN

Seqencia N e C terminais DNA RNAs (mRNA, rRNA e tRNA) Estruturas: 1a, 2a, 3a (4a) Poliamidas

5 3 RNA http://www.clcbio.com/index.php?id=493 advanced primer design tools

Escreva a sequncia dos dois iniciadores de 15 resduos que poderiam ser utilizados para amplificar o seguinte segmento de DNA por PCR:

atggcggcggcggaagaagaaccgaaaccgaaaaaactgaaagtggaagcgccgcaggcgctgcg cgaaaacattctgtttggcatgggcaacccgctgctggatattagcgcggtggtggataaagattttctggata aatatagcctgaaaccgaacgatcagattctggcggaagataacataaagaactgtttgatgaactggtga aaaaatttaaagtggaatatcatgcgggcggcagcacccagaacagcattaaagtggcgcagtggatga ttcagcagccgcataaagcggcgaccttttttggctgcattggcattgataaatttggcgaaattctgaaacgc aaagcggcggaagcgcatgtggatgcgcattattatgaacagaacgaacagccgaccggcacctgc

ATGGCGGCGGCGGAAGAAGAACCGAAACCGAAAAAACTGAAAGTGGAAGCGCCGCAG GCGCTGCGCGAAAACATTCTGTTTGGCATGGGCAACCCGCTGCTGGATATTAGCGCGG TGGTGGATAAAGATTTTCTGGATAAATATAGCCTGAAACCGAACGATCAGATTCTGGCG GAAGATAACATAAAGAACTGTTTGATGAACTGGTGAAAAAATTTAAAGTGGAATATCATG CGGGCGGCAGCACCCAGAACAGCATTAAAGTGGCGCAGTGGATGATTCAGCAGCCGCA TAAAGCGGCGACCTTTTTTGGCTGCATTGGCATTGATAAATTTGGCGAAATTCTGAAACG CAAAGCGGCGGAAGCGCATGTGGATGCGCATTATTATGAACAGAACGAACAGCCGACC GGCACCTGCGCGGCGTGCATTACCGGCGATAACCGCAGCCTGATTGCGAACCTGGCGG CGGCGAACTGCTATAAAAAAGAAAAACATCTGGATCTGGAAAAAAACTGGATGCTGGTG GAAAAAGCGCGCTGTGCTATATTGCGGGCTTTTTTCTGACCGTGAGCCCGGAAAGCGTG CTGAAAGTGGCGCATCATGCGAGCGAAAACAACCGCATTTTTACCCTGAACCTGAGCGC GCCGTTTATTAGCCAGTTTTATAAAGAAAGCCTGATGAAAGTGATGCCGTATGTGGATAT TCTGTTTGGCAACGAAACCGAAGCGGCGACCTTTGCGCGCGAACAGGGCTTTGAAACC AAAGATATTAAAGAAATTGCGAAAAAAACCCAGGCGCTGCCGAAAATGAACAGCAAACG CCAGCGCATTGTGATTTTTACCCAGGGCCGCGATGATACCATTATGGCGACCGAAAGCG AAGTGACCGCGTTTGCGGTGCTGGATCAGGATCAGAAAGAAATTATTGATACCAACGGC GCGGGCGATGCGTTTGTGGGCGGCTTTCTGAGCCAGCTGGTGAGCGATAAACCGCTGA CCGAATGCATTCGCGCGGGCCATTATGCGGCGAGCATTATTATTCGCCGCACCGGCTG CACCTTTCCGGAAAAACCGGATTTTCAT

 a. b. c. d. e. f.

Genoma e proteoma escolher um gene replicar o fragmento de DNA PCR cortar DNA em posio definida ligar com vetor de clonagem inserir o DNA-recombinante numa clula competente selecionar as bactrias com DNA modificado

1. Escolher um gene

2. Desenhar iniciador (primer)

3. Reao de PCR

4. Isolar e purificar fragmento de DNA de tamanho esperado

Gel de agarose eletroforese de DNA

5. Utilizar endonuclease de restrio

6. Ligar o fragmento preparado num vetor de clonagem

7. E. coli cepas diferentes como clulas hospedeiras

Endonucleasa de restrio

http://bcs.whfreeman.com/lodish5e Lodish, H.; et al.; Molecular Cell Biology CD-ROM, 3rd Ed., Freeman, 1996. http://bcs.whfreeman.com/lehninger Nelson, D.; Cox, M.; Lehninger Principles of Biochemistry, 4th Ed., Freeman, 2005. Campbell, M. K.; Lue, R.; Fixen, W.; Purves, W. K.; Understand Biochemistry, Interactive Learning, CD-ROM,Version 1.1, 2000. Voet, D.;Voet, J. G.; Pratt, C. W.; Fundamentos de Bioqumica, Porto Alegre, 2000.