[Construction of nsdAmgh gene disruption mutant in Strempomyces roseoflavus Men-myco-93-63].
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.
Fengying
F
Shen
Weigang
W
Wu
Yanjie
Y
Zhang
Hongda
H
Kou
Hongliu
H
Ji
Yaning
Y
Li
Daqun
D
Liu
D005798:Genes, Bacterial; D005822:Genetic Vectors; D016254:Mutagenesis, Insertional; D016133:Polymerase Chain Reaction; D013302:Streptomyces / Q000235:genetics*