The refinement of species affiliation of some click beetles (Coleoptera: Elateridae) based on the molecular genetic analysis

The article is devoted to data on the approbation of modern methods and the refinement of species affiliation of soil-inhabiting larvae of click beetles (wireworm) based on the molecular genetic analysis (DNAbarcoding). For the first time, data obtained on the complete identity of certain DNA sequences of a number of species of click beetles living in the Kostanay Region (Kazakhstan) — dangerous pests of agricultural crops. At the same time, the World genetic bank (GenBank) did not find identical DNA sequences of decoded DNA nucleotide sequences for a number of studied specimens. The basis for this study was the materials collected in 2018 in the subzone of ordinary black earth on sandy loam soils (Mendykarinsky district). The selection of larvae was carried out by the method of standard soil-zoological samples. The fixation and storage of the selected click beetles larvae was carried out according to the method of preparing samples for molecular genetic analysis with fixation in 96 % alcohol. After the classic identification of the taxonomic position of the collected specimens, the species were identified by genetic analysis on the nucleotide sequence of the cytochrome C oxidase I subunit gene (COI). The assembly and decoding of the DNA nucleotide sequences of the studied samples were carried out using the programs «Codon Code Aligner» and «MEGA-X». As a result of the work carried out in the DNA laboratory of the Museum of Natural History (University of Oslo, Norway), it was possible to identify the complete identity of the DNA sequences of several mass species of click beetles, while a number of decoded DNA sequences of model specimens were absent in the genetic bank, which requires replenishment in it with new data.

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Molecular genetic analysis of chromosome arm 17p and chromosome arm 22q DNA sequences in sporadic pediatric ependymomas

Genes Chromosomes and Cancer ◽

10.1002/(sici)1098-2264(199609)17:1<37::aid-gcc6>3.0.co;2-3 ◽

1996 ◽

Vol 17 (1) ◽

pp. 37-44 ◽

Cited By ~ 70

Author(s):

Markus S. von Haken ◽

Eileen C. White ◽

Laleh Daneshvar-Shyesther ◽

Sharon Sih ◽

Eugene Choi ◽

...

Keyword(s):

Genetic Analysis ◽

Dna Sequences ◽

Molecular Genetic ◽

Molecular Genetic Analysis ◽

Chromosome Arm

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USER Friendly Cloning Coupled with Chitin-Based Natural Transformation Enables Rapid Mutagenesis of Vibrio vulnificus

Applied and Environmental Microbiology ◽

10.1128/aem.02564-08 ◽

2009 ◽

Vol 75 (15) ◽

pp. 4936-4949 ◽

Cited By ~ 65

Author(s):

Paul A. Gulig ◽

Matthew S. Tucker ◽

Patrick C. Thiaville ◽

Jennifer L. Joseph ◽

Roslyn N. Brown

Keyword(s):

Genetic Analysis ◽

Dna Sequences ◽

Vibrio Vulnificus ◽

Molecular Genetic ◽

Opportunistic Pathogen ◽

Molecular Genetic Analysis ◽

Single Step ◽

Expression Vectors ◽

Allelic Exchange ◽

User Friendly

ABSTRACT Vibrio vulnificus is a bacterial contaminant of shellfish and causes highly lethal sepsis and destructive wound infections. A definitive identification of virulence factors using the molecular version of Koch's postulates has been hindered because of difficulties in performing molecular genetic analysis of this opportunistic pathogen. For example, conjugation is required to introduce plasmid DNA, and allelic exchange suicide vectors that rely on sucrose sensitivity for counterselection are not efficient. We therefore incorporated USER friendly cloning techniques into pCVD442-based allelic exchange suicide vectors and other expression vectors to enable the rapid and efficient capture of PCR amplicons. Upstream and downstream DNA sequences flanking genes targeted for deletion were cloned together in a single step. Based on results from Vibrio cholerae, we determined that V. vulnificus becomes naturally transformable with linear DNA during growth on chitin in the form of crab shells. By combining USER friendly cloning and chitin-based transformation, we rapidly and efficiently produced targeted deletions in V. vulnificus, bypassing the need for two-step, suicide vector-mediated allelic exchange. These methods were used to examine the roles of two flagellin loci (flaCDE and flaFBA), the motAB genes, and the cheY-3 gene in motility and to create deletions of rtxC, rtxA1, and fadR. Additionally, chitin-based transformation was useful in moving antibiotic resistance-labeled mutations between V. vulnificus strains by simply coculturing the strains on crab shells. The methods and genetic tools that we developed should be of general use to those performing molecular genetic analysis and manipulation of other gram-negative bacteria.

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Molecular genetic analysis of stygobiotic shrimps of the genus Xiphocaridinella (Crustacea: Decapoda: Atyidae) reveals a connection between distant caves in Central Abkhazia, southwestern Caucasus

International Journal of Speleology ◽

10.5038/1827-806x.50.3.2378 ◽

2021 ◽

Vol 50 (3) ◽

pp. 301-311

Author(s):

Ivan Marin ◽

Ilya Turbanov

Keyword(s):

New Species ◽

Genetic Analysis ◽

Cytochrome Oxidase ◽

Dna Sequences ◽

Genetic Study ◽

Molecular Genetic ◽

Molecular Genetic Analysis ◽

Cytochrome Oxidase I ◽

Mitochondrial Cytochrome ◽

Mitochondrial Cytochrome Oxidase

Based on the morpho-genetic study of stygobiotic shrimps from the genus Xiphocaridinella Sadowsky, 1930 (Crustacea: Decapoda: Atyidae), a hydrogeological connection of a number of distant caves in Central Abkhazia of the southwestern Caucasus is satisfied, which indicates the possibility of using biospeleological studies in some cases to identify karst hydrosystems together with traditional hydrogeological methods. Moreover, a new stygobiotic atyid shrimp from the genus Xiphocaridinella, X. kelasuri sp. n., is described based on morphology and analysis of mitochondrial cytochrome oxidase I DNA sequences from three distant caves. The new species is genetically divergent from relatives and phylogenetically related to Xiphocaridinella smirnovi Marin, 2020, described from the Besletka (=Tskaro) Cave. Recently, the number of described speciesof the genus Xiphocaridinella from Caucasus has increased to 13 species, while the diversity of Xiphocaridinella found in the Besletka (=Tskaro) Cave is increasing to 3 species, which is higher than in any other known cave where Troglocaris-like shrimps have been discovered.

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