scholarly journals HPA-10wb (Laa): Genetic Determination of a New Platelet-Specific Alloantigen on Glycoprotein IIIa and Its Expression in COS-7 Cells

Blood ◽  
1997 ◽  
Vol 89 (7) ◽  
pp. 2422-2428 ◽  
Author(s):  
O. Peyruchaud ◽  
F. Bourre ◽  
M.-C. Morel-Kopp ◽  
D. Reviron ◽  
P. Mercier ◽  
...  
Keyword(s):  
Platelet Destruction ◽  
Genetic Determination ◽  
Fibrinogen Receptor ◽  
Glycoprotein Iiia ◽  
Polymerase Chain ◽  
Heterodimeric Complex ◽  
Alloimmune Thrombocytopenia ◽  
Conformation Polymorphism

Abstract The heterodimeric complex glycoprotein (GP)IIb-IIIa, the fibrinogen receptor of platelets, carries numerous alloantigen systems. These polymorphisms are responsible for the immune response after transfusion or during pregnancy. In the latter case, the mother develops an antibody against an epitope present on fetal platelets, and this results in platelet destruction in the fetus. In this report, we describe the molecular characterization of a new alloantigen (Laa) on GPIIIa responsible for neonatal alloimmune thrombocytopenia (NAIT). Using polymerase chain reaction (PCR)–singlestrand conformation polymorphism (SSCP) and DNA sequencing, we found a point mutation (G to A) in a heterozygous state on the GPIIIa gene leading to amino acid substitution Arg to Gln at position 62 of the mature protein. Transient expression of GPIIb-IIIa complexes in Cos-7 cells using wild-type or mutated GPIIIa cDNA allowed us to demonstrate that this mutation was responsible for expression of the Laa epitope.

scholarly journals First Isolation and Molecular Typing of Pathogenic and Intermediate Leptospira Species from Urine of Symptomatic Dogs

2021 ◽  
Vol 8 (12) ◽  
pp. 304
Author(s):  
Ivana Piredda ◽  
Loris Bertoldi ◽  
Giuseppe Benvenuto ◽  
Bruna Palmas ◽  
Aureliana Pedditzi ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Zoonotic Risk ◽  
Leptospira Spp ◽  
Blood And Urine Samples ◽  
Healthy Dogs ◽  
Polymerase Chain ◽  
Sequence Types ◽  
Leptospira Species

Aim of this study was to evaluate, the presence and diversity of Leptospira spp. in blood and urine samples collected from 175 owned-dogs from Sardinia, Italy. After determination of leptospiral infection by microscopic agglutination test (MAT), urine from MAT-positive dogs were examined by real-time polymerase chain reaction (lipL32 rt-PCR) and then isolated by culture. In order to characterize obtained serovars, positive cultures were then subjected to 16S rRNA and secY sequencing, phylogenetic analysis and Multilocus Sequence Typing (MLST). Results showed that seven dogs (4%; 95% CI: 0–55) had Leptospira DNAs in their urine and five strains were isolated from urine cultures. The three different sequence types (ST17, ST198 and ST24) belonging to Leptospira interrogans genomospecies identified by MLST analyses in this study, confirmed that the leptospiral infection was widespread in Sardinian dogs. We also reported the first characterization of a new Leptospira spp. isolated from urine of one dog living in the study area. Whole genome sequencing and phylogenetic analysis, confirmed that this genospecies was closely related to Leptospira hovindhougenii, an intermediate Leptospira spp. with unknown pathogenicity previously isolated from a rat in Denmark. Further studies are required to clarify whether healthy dogs that shed leptospires in their urine could represent a zoonotic risk for humans in this region.

scholarly journals Sex Determination During Inflorescence Bud Differentiation in Monoecious Pistacia chinensis Bunge

Forests ◽  
2019 ◽  
Vol 10 (3) ◽  
pp. 202 ◽  
Author(s):  
Qian Bai ◽  
Chenyi Zhu ◽  
Xia Lei ◽  
Tao Cao ◽  
Shuchai Su ◽  
...  
Keyword(s):  
Sex Determination ◽  
Early Stage ◽  
Sex Differentiation ◽  
Banding Patterns ◽  
Vegetative Period ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Sex Determination Mechanisms

Pistacia chinensis Bunge is widely acknowledged to be dioecious, but rare monoecious individuals have been found. However, the origin of monoecism and the sex differentiation of different sex types remain intriguing questions. Here, sex expressions were explored by identification of sex-associated DNA markers, determination of the sex stability after grafting, and histological characterization of inflorescence bud development using anatomical analysis. The results showed that (1) although polymorphisms among individuals existed, the banding patterns of Polymerase Chain Reaction (PCR) products for different sex types on the same monoecious tree were consistent; (2) the sex expressions of grafted trees were not consistent with those of scions, indicating that monoecism probably did not originate from a stable bud mutation; and (3) both males and females underwent a bisexual period, then the stamen primordia in female buds degenerated into the second round tepals, while the pistil primordia in male buds gradually disappeared. During the sex differentiation phase, female buds were spindle-shaped, while the male buds were full teardrop-shaped, and male buds were bigger than female buds. Taken together, no sex-associated DNA marker was found, sex expressions were unstable after grafting, and the alternative sex organs appeared in the early stage of sex differentiation, suggesting that sex determination occurred during floral development instead of the early vegetative period. These results indicated that the sex expressions may be affected by environmental factors, increasing the understanding of sex determination mechanisms in P. chinensis and other species.

Étude du polymorphisme enzymatique d'une population de pleurotes des ombellifères : Pleurotus eryngii (Tournus, France)

1983 ◽  
Vol 61 (2) ◽  
pp. 458-465 ◽  
Author(s):  
Marie-Catherine Boisselier-Dubayle
Keyword(s):  
Analytical Method ◽  
Pleurotus Eryngii ◽  
Genetic Determination ◽  
Phosphatase Activities ◽  
Electrophoretic Patterns ◽  
Convenient Means ◽  
Electrophoretic Separations ◽  
Taxonomic Studies

Electrophoretic separations followed by revelations of esterase, aminopeptidase, and phosphatase activities were performed on a population of Pleurotus eryngii (Tournus: Saône valley, France). Electrophoresis on controlled dicaryotic mycelia and their haploid components was used as a convenient means to study the genetic determination of some esterase and phosphatase isoenzymes. The electrophoretic patterns obtained from original isolates (wild dicaryons) show little variability, but internal variations are revealed. These variations allowed for a characterization of four genotypically different individual groups. This analytical method seems to give some interesting data, useful for taxonomic studies.

Genetic determination of Saccharomyces cerevisiae et Saccharomyces bayanus species by PCR/RFLP analysis of the MET2 gene

OENO One ◽  
1996 ◽  
Vol 30 (1) ◽  
pp. 15
Author(s):  
Isabelle Masneuf-Pomarède ◽  
Michel Aigle ◽  
Denis Dubourdieu
Keyword(s):  
Dna Hybridization ◽  
Rflp Analysis ◽  
Distinct Species ◽  
Genetic Determination ◽  
Yeast Strains ◽  
Saccharomyces Bayanus ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Em Gene

<p style="text-align: justify;">Several yeast strains of the species <em>S. cerevisiae</em>, <em>S. bayanus</em> and <em>S. paradoxus</em>, first identified by hybridization experiments and DNA/DNA hybridization were characterized by using Polymerase Chain Reaction/Restriction Fragment Length Polymorphism (PCR/RFLP) of the <em>MET2 gene</em>. The concordance between this tool and classical genetic analyses did not reveal any exception for all the strains analysed, so PCR/RFLP of the MET2 gene proves to be a reliable and fast tool for delimiting <em>S. cerevisiae</em> and <em>S. bayanus</em>. OEnological strains race <em>bayanus</em>, <em>chevalieri</em>, <em>capensis</em> gave <em>S. cerevisiae</em> restriction patterns, whereas most of strains race <em>S. uvarum</em> belong to <em>S. bayanus</em> and displayed a specific chromosomal band patterns different from band patterns of <em>S. cerevisiae</em> strains. To avoid confusion in oenological terminology, oenologists should no longer use the name of <em>bayanus</em> to designate industrial or wild <em>S. cerevisiae</em> Gal- strains, and should consider <em>S. bayanus</em> as a distinct species.</p>

scholarly journals Characterization of a transgenic mouse line over-expressing the human ornithine decarboxylase gene

1991 ◽  
Vol 278 (3) ◽  
pp. 895-898 ◽  
Author(s):  
M Halmekytö ◽  
L Alhonen ◽  
J Wahlfors ◽  
R Sinervirta ◽  
T Eloranta ◽  
...  
Keyword(s):  
Transgenic Mouse ◽  
Ornithine Decarboxylase ◽  
Transgenic Animals ◽  
Metabolizing Enzymes ◽  
Transgenic Mouse Line ◽  
Adenosylmethionine Decarboxylase ◽  
Transgenic Lines ◽  
Polymerase Chain

We have produced several transgenic mouse lines over-expressing the human ornithine decarboxylase (ODC) gene. We have now characterized one of the transgenic lines as regards the tissue accumulation of the polyamines and the activities of their metabolizing enzymes. Among the tissues analysed, the polyamine pattern was most strikingly changed in testis and brain of the transgenic animals. ODC activity was greatly enhanced in all tissues, except kidney, of the transgenic animals. The most dramatic increase, 80-fold, was found in brain of the transgenic mice. The activities of S-adenosylmethionine decarboxylase and spermidine and spermine syntheses were likewise significantly increased in testis of the transgenic animals. The activities of the enzymes involved in the back-conversion of the polyamines, namely spermidine/spermine acetyltransferase and polyamine oxidase, were similar in the transgenic and non-transgenic animals. As analysed by reverse transcriptase/polymerase chain reaction, all the six tissues of the transgenic animals expressed human-specific ODC mRNA. Determination of the half-life of testicular ODC revealed a stabilization of the enzyme in the transgenic males.

scholarly journals Differences in Virulence and Genomic Features of Strains of ‘Candidatus Phytoplasma mali’, the Apple Proliferation Agent

2007 ◽  
Vol 97 (8) ◽  
pp. 964-970 ◽  
Author(s):  
Erich Seemüller ◽  
Bernd Schneider
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Blot Hybridization ◽  
Pcr Amplification ◽  
Restriction Enzymes ◽  
Southern Blot Hybridization ◽  
Apple Proliferation ◽  
Candidatus Phytoplasma Mali ◽  
Polymerase Chain

Root and shoot samples from 24 symptomatic or nonsymptomatic apple trees infected with ‘Candidatus Phytoplasma mali’ were collected at different locations in Germany and France and used to inoculate rootstock M11 top grafted with cv. Golden Delicious. Inoculated trees were monitored over a 12-year period for apple proliferation (AP) symptoms and categorized as not or slightly, moderately, or severely affected. Based on symptomatology, the phytoplasma strains were defined as being avirulent to mildly, moderately, or highly virulent. Determination of phytoplasma titers by quantitative polymerase chain reaction (PCR) with DNA from roots revealed similar phytoplasma concentrations in all virulence groups. Molecular characterization of the strains by differential PCR amplification with five sets of primers resulted in 13 profiles. Six strains that were maintained in periwinkle and tobacco were molecularly characterized in more detail. The genome sizes of these strains as determined by pulsed-field gel electrophoresis using yeast chromosomes as size references ranged between 640 and 680 kb. Cleavage of the chromosome with the rare cutting restriction enzymes ApaI, BamHI, BssHII, MluI, and SmaI resulted in macro fragment patterns distinctly different in all strains. Similar results were obtained by Southern blot hybridization with three probes derived from strain AT. Differential PCR amplification at an annealing temperature of 52°C using eight primer pairs derived from strain AT revealed heterogeneity of target sequences among all strains. Based on these results, there is considerable variability in virulence and genomic traits in ‘Ca. P. mali’. However, correlations between molecular markers and virulence or phytoplasma titer could not be identified.

scholarly journals Characterization of Aggregatibacter actinomycetemcomitans Serotype b Strains with Five Different, Including Two Novel, Leukotoxin Promoter Structures

Vaccines ◽  
2020 ◽  
Vol 8 (3) ◽  
pp. 398 ◽  
Author(s):  
Rolf Claesson ◽  
Huei-Min Chiang ◽  
Mark Lindholm ◽  
Carola Höglund Åberg ◽  
Dorte Haubek ◽  
...  
Keyword(s):  
Genetic Characterization ◽  
Close Association ◽  
Typical Feature ◽  
The Past ◽  
Separate Branch ◽  
Enhanced Expression ◽  
Polymerase Chain ◽  
Serotype B

The JP2 genotype of A. actinomycetemcomitans, serotype b has attracted much interest during the past three decades due to its close association with periodontitis in young individuals and the enhanced expression of a leukotoxin (LtxA). A typical feature of this genotype is a 530-base pair (bp) deletion in the ltxCABD promoter region controlling leukotoxin expression. In the present work, we have characterized serotype b strains with four additional promoter types. Two novel types have been recognized, that is, one with a 230-bp deletion and one with a 172-bp duplication. Moreover, a strain with a 640-bp deletion and three strains with a full-length promoter, including the type strain Y4, were included in the present study. The seven strains were characterized by multi locus sequence typing (MLST) and arbitrarily primed polymerase chain reaction (PCR) and assessed for LtxA production. MLST showed that the strains with the non-JP2-like deletions represented distinct monophyletic groups, whereas the JP2 strain, HK1651, represented a separate branch. LtxA production was high in all three strains with a promoter deletion, whereas the other four strains showed significantly lower levels. It can be concluded that the genetic characterization and determination of LtxA production of A. actinomycetemcomitans isolates from individuals with periodontitis can contribute to the identification of novel virulent genotypes of this bacterium.

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