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Showing 3 results for Housekeeping Gene
M. Noofeli , M. Asadipour , R. Yari,
Volume 9, Issue 2 (9-2018)
Abstract
Aims: Bordetella pertussis is a gram negative and obligated aerobic bacterium that causes pertussis disease and it is a specific pathogen in human. Pertussis is an acute respiratory infection and leads to death in infants. The aim of this study was to analyze housekeeping genes in Bordetella pertussis vaccine strains by multilocus sequence typing (MLST).
Materials and Methods: In the present experimental study, 4 samples of 134 and 509 bacterial strains and 2 standard samples of Tohama I, and 18323 were collected. After biochemical tests, the samples were cultured and separated and the genomic purification of DNA was done by Phenol–chloroform technique and analyzed by MLST. After genome sequencing, the analysis was performed by standard software such as Clustalw 2, MEGA 5.04, and DNASIS Max 3. Sequence similarity of 16S rDNA gene nucleotides was performed, using BLAST software with sequences recorded in the GenBank genome database to compare and determine the sequences similarity.
Findings: Regarding the created bands and the sequence of the game, the housekeeping genes in Bordetella pertussis vaccine strains were approved. The results of the PCR reaction for Pgm, Icd, Gly A, and Tyr B genes showed that all specimens have homogenous genes with a molecular weight of 500bp.
Conclusion: Evaluating the housekeeping genes in Bordetella pertussis vaccine strains by MLST vaccine strains (Razi Institute; Iran) correspond with international standard series and no change or deviation has occurred in the studied genes.
Volume 19, Issue 3 (5-2017)
Abstract
In the current study, molecular typing of 50 Erwinia amylovora strains related to different regions in Iran was evaluated using multi-locus sequence analysis and variable number of tandem repeats. In the first assay, phylogenetic tree based on partial sequences of recombinase A, sigma factor S and a heat shock protein GroEL showed significant identity in studied gene sequences. A single nucleotide variation in groEL was determined in IrGh59 strain related to Crataegus spp. from Ghazvin Province. In VNTR analysis, the same fingerprinting profile similar to E. amylovora reference strain ATCC49946 was yielded for tested strains except NBQ1 and MQ1 which may reflect a unique contaminating source for this disease in Iran. In addition, the honey-bee movements with respect to blossom season probably have a considerable role in fire blight unique dispersal in our area. The NBQ1 and MQ1 strains generated different VNTR profiles, isolated from cultivars NeishabourandEsfehan of Cydonia oblonga plant, respectively. No definite assessment can be expressed in this case. However, possible entry of other infection mass from neighboring countries should be determined. Overall, VNTR profile analyses are recommended as a tool to evaluate genetic differences in E. amylovora populations. In addition, employing more strains from different known sources could be assistance to achieve more accurate results about E. amylovora genetic variation and also fire blight distribution patterns.
Volume 21, Issue 4 (7-2019)
Abstract
In the current study, the phenotypic and molecular properties of twenty-five strains obtained from cankerous tissues or leaf necrotic lesions of different stone fruits were evaluated in north-east of Iran . All strains studied were identified as Xanthomonas arboricola pv. pruni (Xap) based on phenotypic assays and confirmed by means of specific PCR at species and pathovar levels. All obtained strains were pathogenic under artificial inoculation and exhibited brittle necrotic spots on plum leaves of cultivar Santa Rosa under lab conditions. Then, the pathogenic Xap strains were subjected to molecular assays. In a phylogenetic tree constructed with gyrB sequences, no polymorphism was observed in this gene and Iranian Xap strains were clustered with the reference one in a separate group. The ERIC, BOX and REP primer sets generated reproducible genomic PCR profiles in tested strains and, based on combined data for all primers, a low genetic diversity among Xap strains was revealed. In order to achieve results that are more accurate, application of Xap strains from all geographical regions of Iran will be needed to prove little polymorphism observed in Xap population. The current contribution is the first report of molecular homogeneity of Xap strains that were collected from northeastern Iran.
