BACKGROUND: Among all common techniques in sitedirectedmutagenesis, λ Red recombinase system has beenwidely used to knock out chromosomal genes in bacteria. In thismethod, there is always the risk of DNA Linear digestion byhost's restriction enzymes that leads to the low frequency ofrecombination. OBJECTIVES:To overcome this, we constructeda recombinant vector to disrupt phoP gene in Salmonella...

background: among all common techniques in sitedirectedmutagenesis, λ red recombinase system has beenwidely used to knock out chromosomal genes in bacteria. in thismethod, there is always the risk of dna linear digestion byhost's restriction enzymes that leads to the low frequency ofrecombination. objectives:to overcome this, we constructeda recombinant vector to disrupt phop gene in salmonella...

Previous studies have shown that an exposure to sub-inhibitory levels of kanamycin induces capsule synthesis which confers antibiotic resistance. To examine the role of spoT and relA genes in capsule synthesis and acquired physiological antibiotic resistance during sub-inhibitory kanamycin treatment, multiple studies were performed but showed inconclusive results. A potential explanation for th...

Site-directed scarless mutagenesis is an essential tool of modern pathogenesis research. We describe an optimized two-step protocol for genome editing in Salmonella enterica serovar Typhimurium to enable multiple sequential mutagenesis steps in a single strain. The system is based on the λ Red recombinase-catalyzed integration of a selectable antibiotics resistance marker followed by replacemen...

Genome engineering methods in E. coli allow for easy to perform manipulations of the chromosome in vivo with the assistance of the λ-Red recombinase system. These methods generally rely on the insertion of an antibiotic resistance cassette followed by removal of the same cassette, resulting in a two-step procedure for genomic manipulations. Here we describe a method and plasmid system that can ...

BACKGROUND: Colibacillosis, caused by different serotypes of avian pathogenic Escherichia coli (APEC), is one of the important diseases in poultry industry. The isolate O78 is the most prevalent serotype of APEC in Iran. One of the APEC virulence factors, increased serum survival (iss) gene, is related to serum resistance. The usual form of colibacillosis in avian is extraintestinal, and serum ...

E. coli is a model platform for engineering microbes, so genetic circuit design and analysis will be greatly facilitated by simple and effective approaches to introduce genetic constructs into the E. coli chromosome at well-characterised loci. We combined the Red recombinase system of bacteriophage λ and Isothermal Gibson Assembly for rapid integration of novel DNA constructs into the E. coli c...