Construction of an iss deleted mutant strain from a native avian pathogenic Escherichia coli O78: K80 and in vitro serum resistance evaluation of mutant
Authors
Abstract:
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 resistance is applied one way by APEC to reach internal organs; hence, it appears that the control of colibacillosis in poultry regarding the deletion of iss and the construction of a serum sensitive APEC strain is beneficial. Additionally, the knowledge about APEC serum resistance could be extended using mutant strains. OBJECTIVES: The present study was an attempt to generate an iss mutant strain from native APEC-O78 strain |c|1378 and to study the level of serum resistance of native APEC-O78 strain c1378 in comparison with its mutant (APEC-O78 strain c1378|D|iss). METHODS: The lambda red recombinase system was utilized to delete iss gene in native APEC-O78 strain c1378. This strain was first transformed with the plasmid pkD46 to introduce the lambda red recombinase system and then the PCR product with sequence homology to the iss gene and a kanamycin resistance marker was transformed into the APEC-O78 strain c1378. Serum sensitivity of mutant and wild type strain was investigated by microtiter test. RESULTS: The generation of mutant was successful and the iss was replaced with kanamycin resistance cassette. Also, it was observed that the mutant was sensitive to serum. However, serum sensitivity of iss deleted mutant was not statistically different from its parents. CONCLUSIONS: Application of lambda red recombination could be a simple and useful technique for production of a precisely defined gene deletion. Also, there may be some genes that compensate the activity of iss gene.
similar resources
construction of an iss deleted mutant strain from a native avian pathogenic escherichia coli o78: k80 and in vitro serum resistance evaluation of mutant
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 ...
full textPhylogenetic Analysis of bor Gene in an Escherichia coli Strain 1378 (O78:K80) Isolated from an Avian Colibacillosis Case in Tehran, Iran
Colibacillosis is known as a fatal bacterial disease resulting in a high level of commercial loss worldwide. This study amid to elucidate the sequence, genetic characteristics, and phylogeny of the bor gene in Escherichia coli (E. coli) strain c1378 (O78:K80) isolated from avian colibacillosis in Iran and develop a rapid and optimal polymerase chain reaction (PCR) mol...
full textaroa gene sequencing of native escherichia coli o78: k80 isolated from avian colibacillosis in iran
aroa is an important gene which produces aromatic amino acids essential for bacterial life. the sequence of this gene was shown to be different in bacteria, so the purpose of this study was sequencing of aroa gene in native e. coli o78: k80 to determine the putative differences between this strain and other e. coli o78: k80 strains in gene bank. pcr was used to amplify aroa gene in native e. co...
full textThe BarA-UvrY two-component system regulates virulence in avian pathogenic Escherichia coli O78:K80:H9.
The BarA-UvrY two-component system (TCS) in Escherichia coli is known to regulate a number of phenotypic traits. Both in vitro and in vivo assays, including the chicken embryo lethality assay, showed that this TCS regulates virulence in avian pathogenic E. coli (APEC) serotype O78:K80:H9. A number of virulence determinants, such as the abilities to adhere, invade, persist within tissues, surviv...
full textIncreased Pho regulon activation correlates with decreased virulence of an avian pathogenic Escherichia coli O78 strain.
Avian pathogenic Escherichia coli (APEC) strains are associated with respiratory infections, septicemia, cellulitis, peritonitis, and other conditions, since colibacillosis manifests in many ways. The Pho regulon is jointly controlled by the two-component regulatory system PhoBR and by the phosphate-specific transport (Pst) system. To determine the specific roles of the PhoBR regulon and the Ps...
full textCharacterization of Stg fimbriae from an avian pathogenic Escherichia coli O78:K80 strain and assessment of their contribution to colonization of the chicken respiratory tract.
In a previous study, ecs-3, a sequence from avian pathogenic Escherichia coli (APEC) O78:K80 strain chi7122, was found to be expressed in vivo in infected chicken tissues. The region encompassing ecs-3 carries a fimbrial gene cluster that is a putative ortholog of the stg fimbrial gene cluster of Salmonella enterica serovar Typhi. This APEC fimbrial gene cluster, which we have termed stg, is a ...
full textMy Resources
Journal title
volume 8 issue 1
pages 1- 8
publication date 2014-04-01
By following a journal you will be notified via email when a new issue of this journal is published.
Hosted on Doprax cloud platform doprax.com
copyright © 2015-2023