RecET recombination system driving chromosomal target gene replacement in Zymomonas mobilis
Electron. j. biotechnol
; 30: 118-124, nov. 2017. tab, ilus, graf
Artigo
em Inglês
| LILACS
| ID: biblio-1021652
Biblioteca responsável:
CL1.1
ABSTRACT
Background:
Zymomonas mobilis is a Gram-negative microaerophilic bacterium with excellent ethanol-producing capabilities. The RecET recombination system provides an efficient tool for direct targeting of genes in the bacterial chromosome by PCR fragments.Results:
The plasmids pSUZM2a-RecET and pSUZM2a-RecE588T were first developed to co-express RecE or RecE588 and RecT for homologous recombination. Thereafter, the PCR fragments of the tetracycline resistance marker gene flanked by 60 bp of adhA (alcohol dehydrogenase I) or adhB (alcohol dehydrogenase II) homologous sequences were electroporated directly into ZM4 cells harboring pSUZM2a-RecET or pSUZM2a-RecE588T. Both adhA and adhB were replaced by the tetracycline resistance gene in ZM4, yielding two mutant strains, Z. mobilis ZM4 ΔadhA and Z. mobilis ZM4 ΔadhB. These two mutants showed varying extent of reduction in ethanol production, biomass generation, and glucose metabolism. Furthermore, enzyme activity of alcohol dehydrogenase II in Z. mobilis ZM4 ΔadhB exhibited a significant reduction compared to that of wild-type ZM4.Conclusion:
This approach provided a simple and useful method for introducing mutations and heterologous genes in the Z. mobilis genome.
Texto completo:
Disponível
Coleções:
Bases de dados internacionais
Base de dados:
LILACS
Assunto principal:
Zymomonas
/
Recombinação Homóloga
Idioma:
Inglês
Revista:
Electron. j. biotechnol
Assunto da revista:
Biotecnologia
Ano de publicação:
2017
Tipo de documento:
Artigo
/
Documento de projeto
País de afiliação:
China
Instituição/País de afiliação:
Ministry of Education/CN