scholarly journals Using the Developmental GeneBicoidto Identify Species of Forensically Important Blowflies (Diptera: Calliphoridae)

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Seong Hwan Park ◽  
Chung Hyun Park ◽  
Yong Zhang ◽  
Huguo Piao ◽  
Ukhee Chung ◽  
...  
Keyword(s):  
Dna Markers ◽  
Forensic Entomology ◽  
Developmental Stages ◽  
Genomic Sequences ◽  
Phormia Regina ◽  
Calliphora Vicina ◽  
Coding Sequences ◽  
Major Subject ◽  
Flanking Sequences ◽  
Intron 2

Identifying species of insects used to estimate postmortem interval (PMI) is a major subject in forensic entomology. Because forensic insect specimens are morphologically uniform and are obtained at various developmental stages, DNA markers are greatly needed. To develop new autosomal DNA markers to identify species, partial genomic sequences of thebicoid(bcd) genes, containing the homeobox and its flanking sequences, from 12 blowfly species (Aldrichina grahami, Calliphora vicina, Calliphora lata, Triceratopyga calliphoroides, Chrysomya megacephala, Chrysomya pinguis, Phormia regina, Lucilia ampullacea, Lucilia caesar, Lucilia illustris, Hemipyrellia ligurriensandLucilia sericata; Calliphoridae: Diptera) were determined and analyzed. This study first sequenced the ten blowfly species other thanC. vicinaandL. sericata. Based on thebcdsequences of these 12 blowfly species, a phylogenetic tree was constructed that discriminates the subfamilies of Calliphoridae (Luciliinae, Chrysomyinae, and Calliphorinae) and most blowfly species. Even partial genomic sequences of about 500 bp can distinguish most blowfly species. The short intron 2 and coding sequences downstream of thebcdhomeobox in exon 3 could be utilized to develop DNA markers for forensic applications. These gene sequences are important in the evolution of insect developmental biology and are potentially useful for identifying insect species in forensic science.

scholarly journals Do longer sequences improve the accuracy of identification of forensically important Calliphoridae species?

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e5962 ◽  
Author(s):  
Sara Bortolini ◽  
Giorgia Giordani ◽  
Fabiola Tuccia ◽  
Lara Maistrello ◽  
Stefano Vanin
Keyword(s):  
Forensic Entomology ◽  
Unsolved Problem ◽  
Phormia Regina ◽  
Calliphora Vicina ◽  
Morphological Identification ◽  
Correct Identification ◽  
Closely Related Species ◽  
Crucial Step ◽  
Post Mortem Interval ◽  
Megaselia Scalaris

Species identification is a crucial step in forensic entomology. In several cases the calculation of the larval age allows the estimation of the minimum Post-Mortem Interval (mPMI). A correct identification of the species is the first step for a correct mPMI estimation. To overcome the difficulties due to the morphological identification especially of the immature stages, a molecular approach can be applied. However, difficulties in separation of closely related species are still an unsolved problem. Sequences of 4 different genes (COI, ND5, EF-1α, PER) of 13 different fly species collected during forensic experiments (Calliphora vicina, Calliphora vomitoria, Lucilia sericata, Lucilia illustris, Lucilia caesar, Chrysomya albiceps, Phormia regina, Cynomya mortuorum, Sarcophaga sp., Hydrotaea sp., Fannia scalaris, Piophila sp., Megaselia scalaris) were evaluated for their capability to identify correctly the species. Three concatenated sequences were obtained combining the four genes in order to verify if longer sequences increase the probability of a correct identification. The obtained results showed that this rule does not work for the species L. caesar and L. illustris. Future works on other DNA regions are suggested to solve this taxonomic issue.

scholarly journals Analysis of HLA-G long-read genomic sequences in mother–offspring pairs with preeclampsia

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Ayako Nishizawa ◽  
Kazuki Kumada ◽  
Keiko Tateno ◽  
Maiko Wagata ◽  
Sakae Saito ◽  
...  
Keyword(s):  
Single Molecule ◽  
Gene Polymorphisms ◽  
Genomic Dna ◽  
Genomic Sequences ◽  
Genomic Sequencing ◽  
Public Database ◽  
Coding Sequences ◽  
Pacbio Rs Ii ◽  
Potential Association ◽  
Long Read

AbstractPreeclampsia is a pregnancy-induced disorder that is characterized by hypertension and is a leading cause of perinatal and maternal–fetal morbidity and mortality. HLA-G is thought to play important roles in maternal–fetal immune tolerance, and the associations between HLA-G gene polymorphisms and the onset of pregnancy-related diseases have been explored extensively. Because contiguous genomic sequencing is difficult, the association between the HLA-G genotype and preeclampsia onset is controversial. In this study, genomic sequences of the HLA-G region (5.2 kb) from 31 pairs of mother–offspring genomic DNA samples (18 pairs from normal pregnancies/births and 13 from preeclampsia births) were obtained by single-molecule real-time sequencing using the PacBio RS II platform. The HLA-G alleles identified in our cohort matched seven known HLA-G alleles, but we also identified two new HLA-G alleles at the fourth-field resolution and compared them with nucleotide sequences from a public database that consisted of coding sequences that cover the 3.1-kb HLA-G gene span. Intriguingly, a potential association between preeclampsia onset and the poly T stretch within the downstream region of the HLA-G*01:01:01:01 allele was found. Our study suggests that long-read sequencing of HLA-G will provide clues for characterizing HLA-G variants that are involved in the pathophysiology of preeclampsia.

scholarly journals Impact of Comingled Heterospecific Assemblages on Developmentally Based Estimates of the Post-Mortem Interval—A Study with Lucilia sericata (Meigen), Phormia regina (Meigen) and Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae)

Insects ◽  
2021 ◽  
Vol 12 (4) ◽  
pp. 280
Author(s):  
Krystal R. Hans ◽  
Sherah L. Vanlaerhoven
Keyword(s):  
Growth Rate ◽  
Development Time ◽  
Lucilia Sericata ◽  
Phormia Regina ◽  
Calliphora Vicina ◽  
Immature Stages ◽  
Post Mortem ◽  
Blow Fly ◽  
Blow Flies ◽  
Post Mortem Interval

Estimates of the minimum post-mortem interval (mPMI) using the development rate of blow flies (Diptera: Calliphoridae) are common in modern forensic entomology casework. These estimates are based on single species developing in the absence of heterospecific interactions. Yet, in real-world situations, it is not uncommon to have 2 or more blow fly species developing on a body. Species interactions have the potential to change the acceptance of resources as suitable for oviposition, the timing of oviposition, growth rate, size and development time of immature stages, as well as impacting the survival of immature stages to reach adult. This study measured larval development and growth rate of the blow flies Lucilia sericata (Meigen, 1826), Phormia regina (Meigen, 1826) and Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae) over five constant temperatures (15, 20, 25, 30, 35 °C), in the presence of conspecifics or two-species heterospecific assemblages. Temperature and species treatment interacted such that L. sericata larvae gained mass more rapidly when in the presence of P. regina at 20 and 30 °C, however only developed faster at first instar. At later stages, the presence of P. regina slowed development of L. sericata immatures. Development time of C. vicina immatures was not affected by the presence of P. regina, however larvae gained mass more slowly. Development time of P. regina immatures was faster in the presence of either L. sericata or C. vicina until third instar, at which point, the presence of L. sericata was neutral whereas C. vicina negatively impacted development time. Phormia regina larvae gained mass more rapidly in the presence of L. sericata at 20 °C but were negatively impacted at 25 °C by the presence of either L. sericata or C. vicina. The results of this study indicate that metrics such as development time or larval mass used for estimating mPMI with blow flies are impacted by the presence of comingled heterospecific blow fly assemblages. As the effects of heterospecific assemblages are not uniformly positive or negative between stages, temperatures or species combinations, more research into these effects is vital. Until then, caution should be used when estimating mPMI in cases with multiple blow fly species interacting on a body.

A and T homopolymeric stretches mediate a DNA inversion in Plasmodium falciparum which results in loss of gene expression

1990 ◽  
Vol 10 (6) ◽  
pp. 3243-3246
Author(s):  
L G Pologe ◽  
D de Bruin ◽  
J V Ravetch
Keyword(s):  
Gene Expression ◽  
Plasmodium Falciparum ◽  
Gene Structure ◽  
Infected Erythrocyte ◽  
Dna Rearrangement ◽  
Coding Sequences ◽  
Dna Inversion ◽  
Erythrocyte Surface ◽  
Stable Substrate ◽  
Flanking Sequences

Ring-infected erythrocyte surface antigen-negative isolates of Plasmodium falciparum demonstrate a complex DNA rearrangement with inversion of 5' coding sequences, deletion of upstream and flanking sequences, and healing of the truncated chromosome by telomere addition. An inversion intermediate that results in the telomeric gene structure for RESA has been identified in the pathway. This inversion creates a mitotically stable substrate for the sequence-specific addition of telomere repeats at the deletion breakpoint.

scholarly journals A and T homopolymeric stretches mediate a DNA inversion in Plasmodium falciparum which results in loss of gene expression.

1990 ◽  
Vol 10 (6) ◽  
pp. 3243-3246 ◽  
Author(s):  
L G Pologe ◽  
D de Bruin ◽  
J V Ravetch
Keyword(s):  
Gene Expression ◽  
Plasmodium Falciparum ◽  
Gene Structure ◽  
Infected Erythrocyte ◽  
Dna Rearrangement ◽  
Coding Sequences ◽  
Dna Inversion ◽  
Erythrocyte Surface ◽  
Stable Substrate ◽  
Flanking Sequences

Ring-infected erythrocyte surface antigen-negative isolates of Plasmodium falciparum demonstrate a complex DNA rearrangement with inversion of 5' coding sequences, deletion of upstream and flanking sequences, and healing of the truncated chromosome by telomere addition. An inversion intermediate that results in the telomeric gene structure for RESA has been identified in the pathway. This inversion creates a mitotically stable substrate for the sequence-specific addition of telomere repeats at the deletion breakpoint.

scholarly journals First record ofPhormia regina(Meigen, 1826) (Diptera: Calliphoridae) from mummies at the Sant’Antonio Abate Cathedral of Castelsardo, Sardinia, Italy

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e4176 ◽  
Author(s):  
Giorgia Giordani ◽  
Fabiola Tuccia ◽  
Ignazio Floris ◽  
Stefano Vanin
Keyword(s):  
Warm Season ◽  
First Record ◽  
Phormia Regina ◽  
Calliphora Vicina ◽  
Single Specimen ◽  
The Past ◽  
Human Decomposition ◽  
The Family ◽  
The Moment ◽  
Biodiversity Changes

The studies of insects from archaeological contexts can provide an important supplement of information to reconstruct past events, climate and environments. Furthermore, the list of the species present in an area in the past allows the reconstruction of the entomofauna on that area at that time, that can be different from the nowadays condition, providing information about biodiversity changes. In this work, the results of a funerary archaeoentomological study on samples collected from mummified corpses discovered during the restoration of the crypt of the Sant’Antonio Abate Cathedral of Castelsardo (Sardinia, Italy) are reported. The majority of the sampled specimens were Diptera puparia, whereas only few Lepidoptera cocoons and some Coleoptera fragments were isolated. Among Diptera, Calliphoridae puparia were identified asPhormia regina(Meigen, 1826) andCalliphora vicina, (Robineau-Desvoidy, 1830) both species typical of the first colonization waves of exposed bodies. Three puparia fragments were also identified as belonging to aSarcophagaMeigen, 1826, species (Sarcophagidae). Several Muscidae puparia of the speciesHydrotaea capensis(Weidmermann, 1818), a late colonizer of bodies, and typical of buried bodies were also collected. The few moth (Lepidoptera) cocoons were identified as belonging to the family Tineidae. This family comprises species feeding on dry tissues and hair typical of the later phases of the human decomposition. Among Coleoptera a single specimen in the family Histeridae,Saprinus semistriatus(Scriba, 1790) and a single elytra, potentially of a species in the family Tenebrionidae, were also collected. Overall, the samples collected indicated an initial colonization of the bodies in an exposed context, mainly in a warm season. This research allows the finding of elements indicating the presence, at least in the past, ofP. reginain Sardinia. This species at the moment seems extinct from Sardinia while it is quite common in the continent.

scholarly journals Application of Serial Analysis of Gene Expression to the Study of the Gene Expression Profile ofLeishmania infantum chagasiPromastigote

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Adelino Soares Lima Neto ◽  
Osvaldo Pompílio de Melo Neto ◽  
Carlos Henrique Nery Costa
Keyword(s):  
Gene Expression ◽  
Life Cycle ◽  
Gene Expression Profile ◽  
Expression Profile ◽  
Leishmania Infantum ◽  
Open Reading Frames ◽  
Genomic Sequences ◽  
Coding Sequences ◽  
Key Genes ◽  
Reading Frames

This study describes the application of the LongSAGE methodology to study the gene expression profile in promastigotes ofLeishmania infantum chagasi. A tag library was created using the LongSAGE method and consisted of 14,208 tags of 17 bases. Of these, 8,427 (59.3%) were distinct. BLAST research of the 1,645 most abundant tags showed that 12.8% of them identified the coding sequences of genes, while 82% (1,349/1,645) identified one or more genomic sequences that did not correspond with open reading frames. Only 5.2% (84/1,645) of the tags were not aligned to any position in theL. infantum genome. The UTR size ofLeishmaniaand the lack of CATG sites in some transcripts were decisive for the generation of tags in these regions. Additional analysis will allow a better understanding of the expression profile and discovering the key genes in this life cycle.

Stephanoprora amurensis sp. nov., Echinochasmus milvi Yamaguti, 1939 and E. suifunensis Besprozvannykh, 1991 from the Russian southern Far East and their phylogenetic relationships within the Echinochasmidae Odhner 1910

Parasitology ◽  
2020 ◽  
Vol 147 (13) ◽  
pp. 1469-1479
Author(s):  
Y. V. Tatonova ◽  
A. V. Izrailskaia ◽  
V. V. Besprozvannykh
Keyword(s):  
Dna Markers ◽  
Phylogenetic Relationships ◽  
Developmental Stages ◽  
Far East ◽  
Genetic Data ◽  
Freshwater Snails ◽  
Its2 Region ◽  
Rrna Gene ◽  
28S Rrna ◽  
Southern Far East

AbstractMature worms of Stephanoprora amurensis sp. nov. were obtained in an experimental study of its life cycle. In the Russian southern Far East, this trematode circulates using freshwater snails Parajuga subtegulata, freshwater fish and birds as the first, second intermediate and final hosts, respectively. Stephanoprora amurensis sp. nov. differs from the well-known representatives of Stephanoprora in a number of morphometric indicators of the developmental stages. The validity of the species was also confirmed by nuclear and mitochondrial DNA markers. In addition, new genetic data were obtained for Echinochasmus suifunensis and Echinochasmus milvi. An analysis of phylogenetic relationships within Echinochasmidae based on the 28S rRNA gene and ITS2 region identified two clusters, one of which combines species of Echinochasmus with 20–22 collar spines and short-tailed cercariae, and the other which includes Stephanoprora spp. and a number of representatives of Echinochasmus with 24 collar spines and long-tailed cercariae. The results of phylogenetic analysis based on ITS2 data show interfamily level of differences between the two clusters and intergeneric differentiation between the three subclusters uniting the species of Stephanoprora and Echinochasmus.

Locus control region activity by 5′HS3 requires a functional interaction with β-globin gene regulatory elements: expression of novel β/γ-globin hybrid transgenes

Blood ◽  
2000 ◽  
Vol 95 (10) ◽  
pp. 3242-3249 ◽  
Author(s):  
Joel E. Rubin ◽  
Peter Pasceri ◽  
Xiumei Wu ◽  
Philippe Leboulch ◽  
James Ellis
Keyword(s):  
Gene Therapy ◽  
Transgenic Mice ◽  
Control Region ◽  
Globin Gene ◽  
Single Copy ◽  
Gene Sequences ◽  
Coding Sequences ◽  
Globin Promoter ◽  
Intron 2 ◽  
Chromatin Opening

Abstract The human β-globin locus control region (LCR) contains chromatin opening and transcriptional enhancement activities that are important to include in β-globin gene therapy vectors. We previously used single-copy transgenic mice to map chromatin opening activity to the 5′HS3 LCR element. Here, we test novel hybrid globin genes to identify β-globin gene sequences that functionally interact with 5′HS3. First, we show that an 850-base pair (bp) 5′HS3 element activates high-level β-globin gene expression in fetal livers of 17 of 17 transgenic mice, including 3 single-copy animals, but fails to reproducibly activate Aγ-globin transgenes. To identify the β-globin gene sequences required for LCR activity by 5′HS3, we linked the 815-bp β-globin promoter to Aγ-globin coding sequences (BGT34), together with either the β-globin intron 2 (BGT35), the β-globin 3′ enhancer (BGT54), or both intron 2 and the 3′ enhancer (BGT50). Of these transgenes, only BGT50 reproducibly expresses Aγ-globin RNA (including 7 of 7 single-copy animals, averaging 71% per copy). Modifications to BGT50 show that LCR activity is detected after replacing the β-globin promoter with the 700-bp Aγ-globin promoter, but is abrogated when an AT-rich region is deleted from β-globin intron 2. We conclude that LCR activity by 5′HS3 on globin promoters requires the simultaneous presence of β-globin intron 2 sequences and the 260-bp 3′ β-globin enhancer. The BGT50 construct extends the utility of the 5′HS3 element to include erythroid expression of nonadult β-globin coding sequences in transgenic animals and its ability to express antisickling γ-globin coding sequences at single copy are ideal characteristics for a gene therapy cassette.

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