Construction of nsdAmgh gene disruption mutant in Strempomyces roseoflavus Men-myco-93-63 / 生物工程学报
Chinese Journal of Biotechnology
; (12): 1741-1752, 2015.
Article
in Zh
| WPRIM
| ID: wpr-337461
Responsible library:
WPRO
ABSTRACT
Insertional mutagenesis is a widely used method to determine the function(s) of a gene. To study the function(s) of the gene nsdAmgh in Streptomyces roseoflavus, a homologous recombination vector pSRNA2500 was structured in this paper. The recombination donor vector was then transformed into Strempomyces roseoflavus strain Men-myco-93-63 by conjugative transfer. The transformants were subjected to selection under the pressure of high temperature and appropriate antibiotics. As a result, several disrupted mutants of nsdAmgh gene, with a phenotype of Am(s)Km(r), were isolated and verified using PCR and Dot-blotting and Southern blotting hybridization methods. Functional analysis showed that the disrupted mutants of nsdAmgh had a two-fold higher inhibition against Verticillium dahlia Kleb than that of the wild strain Men-myco-93-63, which all will provide a new study route for future research about positive and negative regulator in Men-myco-93-63.
Full text:
1
Index:
WPRIM
Main subject:
Streptomyces
/
Polymerase Chain Reaction
/
Mutagenesis, Insertional
/
Genes, Bacterial
/
Genetic Vectors
/
Genetics
Language:
Zh
Journal:
Chinese Journal of Biotechnology
Year:
2015
Type:
Article