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Registro Completo |
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Biblioteca(s): |
Embrapa Meio Norte / UEP-Parnaíba. |
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Data corrente: |
28/07/1995 |
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Data da última atualização: |
28/07/1995 |
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Autoria: |
NUGENT, M. E.; PRIMROSE, S. B.; TACON, W. C. A. |
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Afiliação: |
Department of Microbiology and Process Research, Searle Research and Development. |
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Título: |
The stability of recombinant DNA. |
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Ano de publicação: |
0 |
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Fonte/Imprenta: |
s.n.t. |
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Páginas: |
p.271-285 |
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Idioma: |
Inglês |
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Notas: |
Separata. |
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Conteúdo: |
A high copy number mutant of a trp expression plasmid has been used toincrease the expression of the cloned gene urogastrone. This mutation results in up to an eightfold increase in copy number over pBR322. However, when the mutation was cloned into a urogastrone-producing plasmid, only 5% of the resulting recombinants were of the right construction (pMN38-2); 90% had undergone deletions and 5% ha insertions. Attemptwas made to increase urogastrone expression further by cloning a triple trp promoter fragment into pMN38-2. Although there were problems dueto structural instability 10% of recombinants had the correct structure (pMN37-1). However, this plasmid did not lead to any increase in urogastrone production over pMN38-2. In expression studies using there + E. coli K12 HW27 pMN37-1, instability was observed due to homologous recombination between promoter units whereas in the rec strain HB101 drops in copy number were observed. A plasmid, pWT600, was constructed which carries a fusion between a beta-interferon gene and a beta-galactosidase gene. The expression of this gene fusion was under trp promoter control. Cells carrying pWT600 rapidly threw off mutants with decreased expression of this fusion. This was due either to a chromosomal mutation resulting in a 10-fold drop in copy number or to insertions of IS10 from the chromosome into pWT600. From the observations on structural instability, it is important to check the structure and copy number of plasmids during studies designed to maximize expression. MenosA high copy number mutant of a trp expression plasmid has been used toincrease the expression of the cloned gene urogastrone. This mutation results in up to an eightfold increase in copy number over pBR322. However, when the mutation was cloned into a urogastrone-producing plasmid, only 5% of the resulting recombinants were of the right construction (pMN38-2); 90% had undergone deletions and 5% ha insertions. Attemptwas made to increase urogastrone expression further by cloning a triple trp promoter fragment into pMN38-2. Although there were problems dueto structural instability 10% of recombinants had the correct structure (pMN37-1). However, this plasmid did not lead to any increase in urogastrone production over pMN38-2. In expression studies using there + E. coli K12 HW27 pMN37-1, instability was observed due to homologous recombination between promoter units whereas in the rec strain HB101 drops in copy number were observed. A plasmid, pWT600, was constructed which carries a fusion between a beta-interferon gene and a beta-galactosidase gene. The expression of this gene fusion was under trp promoter control. Cells carrying pWT600 rapidly threw off mutants with decreased expression of this fusion. This was due either to a chromosomal mutation resulting in a 10-fold drop in copy number or to insertions of IS10 from the chromosome into pWT600. From the observations on structural instability, it is important to check the structure and copy number of plasmids during studies ... Mostrar Tudo |
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Palavras-Chave: |
Clonagem genica; DNA recombinante; Estabilidade; Expressao plasmidial. |
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Categoria do assunto: |
-- |
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Marc: |
LEADER 02038naa a2200217 a 4500
001 1074997
005 1995-07-28
008 bl --- 0-- u #d
100 1 $aNUGENT, M. E.
245 $aThe stability of recombinant DNA.
260 $c0
300 $ap.271-285
500 $aSeparata.
520 $aA high copy number mutant of a trp expression plasmid has been used toincrease the expression of the cloned gene urogastrone. This mutation results in up to an eightfold increase in copy number over pBR322. However, when the mutation was cloned into a urogastrone-producing plasmid, only 5% of the resulting recombinants were of the right construction (pMN38-2); 90% had undergone deletions and 5% ha insertions. Attemptwas made to increase urogastrone expression further by cloning a triple trp promoter fragment into pMN38-2. Although there were problems dueto structural instability 10% of recombinants had the correct structure (pMN37-1). However, this plasmid did not lead to any increase in urogastrone production over pMN38-2. In expression studies using there + E. coli K12 HW27 pMN37-1, instability was observed due to homologous recombination between promoter units whereas in the rec strain HB101 drops in copy number were observed. A plasmid, pWT600, was constructed which carries a fusion between a beta-interferon gene and a beta-galactosidase gene. The expression of this gene fusion was under trp promoter control. Cells carrying pWT600 rapidly threw off mutants with decreased expression of this fusion. This was due either to a chromosomal mutation resulting in a 10-fold drop in copy number or to insertions of IS10 from the chromosome into pWT600. From the observations on structural instability, it is important to check the structure and copy number of plasmids during studies designed to maximize expression.
653 $aClonagem genica
653 $aDNA recombinante
653 $aEstabilidade
653 $aExpressao plasmidial
700 1 $aPRIMROSE, S. B.
700 1 $aTACON, W. C. A.
773 $ts.n.t.
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Registro original: |
Embrapa Meio Norte / UEP-Parnaíba (CPAMN-UEPP) |
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Biblioteca |
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Registro |
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Voltar
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Registro Completo
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Biblioteca(s): |
Embrapa Tabuleiros Costeiros. |
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Data corrente: |
24/10/2014 |
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Data da última atualização: |
24/10/2014 |
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Tipo da produção científica: |
Artigo em Anais de Congresso |
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Autoria: |
MIRANDA, I. C. D.; OLIVEIRA, S. D. da S. |
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Título: |
Indução de dihaplóides em acessos de coqueiro anão verde do Brasil. |
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Ano de publicação: |
2014 |
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Fonte/Imprenta: |
In: SEMINÁRIO DE INICIAÇÃO CIENTÍFI CA E PÓS-GRADUAÇÃO DA EMBRAPA TABULEIROS COSTEIROS, 4., 2014, Aracaju. Anais... Brasília, DF: Embrapa, 2014. |
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Idioma: |
Português |
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Palavras-Chave: |
Coconut; Coqueiro anão. |
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Thesagro: |
Coco. |
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Categoria do assunto: |
-- |
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URL: |
https://ainfo.cnptia.embrapa.br/digital/bitstream/item/110533/1/256.pdf
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Marc: |
LEADER 00549nam a2200145 a 4500
001 1998315
005 2014-10-24
008 2014 bl uuuu u00u1 u #d
100 1 $aMIRANDA, I. C. D.
245 $aIndução de dihaplóides em acessos de coqueiro anão verde do Brasil.$h[electronic resource]
260 $aIn: SEMINÁRIO DE INICIAÇÃO CIENTÍFI CA E PÓS-GRADUAÇÃO DA EMBRAPA TABULEIROS COSTEIROS, 4., 2014, Aracaju. Anais... Brasília, DF: Embrapa$c2014
650 $aCoco
653 $aCoconut
653 $aCoqueiro anão
700 1 $aOLIVEIRA, S. D. da S.
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Registro original: |
Embrapa Tabuleiros Costeiros (CPATC) |
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