Genetic Diversity of the Flavohemoprotein Gene of Giardia lamblia : Evidence for High Allelic Heterozygosity and Copy Number Variation

The flavohemoprotein (gFlHb) in plays an important role in managing nitrosative and oxidative stress, and potentially also in virulence and nitroimidazole drug tolerance. The aim of this study was to analyze the genetic diversity of in assemblages A and B clinical isolates. genes from 20 cultured cl...

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Bibliographic Details
Published in:Infection and drug resistance 2020-01, Vol.13, p.4531-4545
Main Authors: Saghaug, Christina S, Klotz, Christian, Kallio, Juha P, Aebischer, Toni, Langeland, Nina, Hanevik, Kurt
Format: Article
Language:English
Subjects:
Adaptability
allele
Alleles
Biological diversity
Clinical isolates
Cloning
Comparative analysis
Copy number
copy number variation
Deoxyribonucleic acid
DNA
DNA sequencing
Drug tolerance
Enzymes
Experiments
flavohemoprotein
Genes
Genetic diversity
Genetic research
Genetic variability
Genomes
Genomics
Giardia
Heterozygosity
Homology
Metronidazole
Nitrates
Nitroimidazole
nitrosative stress
Nucleotide sequencing
Original Research
Oxidative stress
Plasmids
Protein structure
Proteins
Structure-function relationships
Toxicity
Virulence
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Summary:The flavohemoprotein (gFlHb) in plays an important role in managing nitrosative and oxidative stress, and potentially also in virulence and nitroimidazole drug tolerance. The aim of this study was to analyze the genetic diversity of in assemblages A and B clinical isolates. genes from 20 cultured clinical isolates were subjected to PCR amplification and cloning, followed by Sanger sequencing. Sequences of all cloned PCR fragments from each isolate were analyzed for single nucleotide variants (SNVs) and compared to genomic Illumina sequence data. Identical clone sequences were sorted into alleles, and diversity was further analyzed. The number of gene copies was assessed by mining PacBio de novo assembled genomes in eight isolates. Homology models for assessment of SNV's potential impact on protein function were created using Phyre2. A variable copy number of the gene, between two and six copies, depending on isolate, was found. A total of 37 distinct sequences, representing different alleles of the gene, were identified in AII isolates, and 41 were identified in B isolates. In some isolates, up to 12 different alleles were found. The total allelic diversity was high for both assemblages (>0.9) and was coupled with a nucleotide diversity of
ISSN:1178-6973
1178-6973
DOI:10.2147/IDR.S274543