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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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| Registros recuperados : 8 | |
| 1. |  | LIMA, T. A. de M. de; LIMA, G. G. de; COSTA, L. N. da; NUGENT, M.; MAGALHAES, W. L. E.; KUROMOTO, N. K. Evaluation of titanium nitride coatings in bandsaw blades for wood splitting by cold plasma. Wood Material Science & Engineering, v. 18, n. 1, p. 130-140, 2023.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: B - 1 |
| Biblioteca(s): Embrapa Florestas. |
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| 2. | | LIMA, G. G. de; CHEE, B. S.; MORITZ, V. F.; CORTESE, Y. J.; MAGALHAES, W. L. E.; DEVINE, D. M.; NUGENT, M. J. D. The production of a novel poly(vinyl alcohol) hydrogel cryogenic spheres for immediate release using a droplet system. Biomedical Physics & Engineering Express, v. 5, n. 4, 045017, Jul. 2019. 13 p.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: B - 3 |
| Biblioteca(s): Embrapa Florestas. |
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| 3. | | LIMA, G. G. de; FERREIRA, B. D.; MATOS, M.; PEREIRA, B. L.; NUGENT, M. J. D.; HANSEL, F. A.; MAGALHAES, W. L. E. Effect of cellulose size-concentration on the structure of polyvinyl alcohol hydrogels. Carbohydrate Polymers, v. 245, 116612, Oct. 2020. 10 p.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: A - 1 |
| Biblioteca(s): Embrapa Florestas. |
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| 4. | | LIMA, G. G. de; BARBOSA, R. R. C.; SANTOS, M. P. de A.; CHEE, B. S.; MAGALHAES, W. L. E.; DEVINE, D. M.; NUGENT, M. J. D. Effect of unidirectional freezing using a thermal camera on polyvinyl (alcohol) for aligned porous cryogels. Quantitative InfraRed Thermography Journal, v. 18, n. 3, p. 177-186, 2021.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: A - 2 |
| Biblioteca(s): Embrapa Florestas. |
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| 5. | | LIMA, G. G. de; ELTER, J. K.; CHEE, B. S.; MAGALHAES, W. L. E.; DEVINE, D. M.; NUGENT, M. J. D.; SÁ, M. J. C. de. A tough and novel dual-response PAA/P(NiPAAM-co-PEGDMA) IPN hydrogels with ceramics by photopolymerization for consolidation of bone fragments following fracture. Biomedical Materials, v. 14, n. 5, Sept. 2019. 14 p.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: A - 1 |
| Biblioteca(s): Embrapa Florestas. |
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| 6. | | ALIABADI, M.; CHEE, B. S.; MATOS, M.; CORTESE, Y. J.; NUGENT, M. J. D.; LIMA, T. A. M. de; MAGALHAES, W. L. E.; LIMA, G. G. de. Yerba mate extract in microfibrillated cellulose and corn starch films as a potential wound healing bandage. Polymers, v. 12, 2807, 2020.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: A - 1 |
| Biblioteca(s): Embrapa Florestas. |
|  |
| 7. | | LIMA, T. A. de M. de; LIMA, G. G. de; CHEE, B. S.; HENN, J. G.; CORTESE, Y. J.; MATOS, M.; HELM, C. V.; MAGALHAES, W. L. E.; NUGENT, M. J. D. Characterization of gels and films produced from pinhão seed coat nanocellulose as a potential use for wound healing dressings and screening of Its compounds towards antitumour effects. Polymers, v. 14, 2776, 2022.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: A - 1 |
| Biblioteca(s): Embrapa Florestas. |
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| 8. | | ALIABADI, M.; CHEE, B. S.; MATOS, M.; CORTESE, Y. J.; NUGENT, M. J. D.; LIMA, T. A. M. de; MAGALHAES, W. L. E.; LIMA, G. G. de; FIROUZABADI, M. D. Microfibrillated cellulose films containing chitosan and tannic acid for wound healing applications. Journal of Materials Science. Materials in Medicine, v. 32, n. 6, article 67, June 2021. 12 p.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: A - 1 |
| Biblioteca(s): Embrapa Florestas. |
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| Registros recuperados : 8 | |
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