Molecular characterization of Ovar-DRB1 exon2 gene in Garole sheep resilient to gastrointestinal nematodes

2015 ◽  
Author(s):  
C. Paswan ◽  
L. L.L. Prince ◽  
R. Kumar ◽  
C. P. Swarnkar ◽  
D. Singh ◽  
...  
Keyword(s):  
Disease Resistance ◽  
Molecular Characterization ◽  
Molecular Mechanisms ◽  
Gastrointestinal Nematodes ◽  
Exon 2 ◽  
Pcr Rflp

Ovine MHC DRB1 exon 2 (Ovar-DRB1.2) gene is arguably one of the critical genes, responsible for disease resistance against parasite in animals. This study was carried out for indepth analysis of polymorphism in MHC DRB1.2 gene and to explore the underlying molecular mechanisms for the development of disease resistance in Garole. PCR-RFLP study revealed that the gene is polymorphic in nature. The frequency of allele ‘A’ for endonuclease SacI and Allele ‘B’ for endonuclease Hin1I were significantly higher in Garole population.

scholarly journals Molecular Characterization of Pathogenicity Locus (PaLoc) and tcdC Genetic Diversity Among tcdA+B+ Clostridioides Difficile Clinical Isolates in Tehran, Iran

2020 ◽  
Author(s):  
Mansoor Kodori ◽  
Zohreh Ghalavand ◽  
Abbas Yadegar ◽  
Gita Eslami ◽  
Masoumeh Azimirad ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Molecular Characterization ◽  
Disease Severity ◽  
Genetic Variants ◽  
Rflp Analysis ◽  
Clostridioides Difficile ◽  
Pcr Rflp ◽  
The Common ◽  
Healthcare Associated

Abstract Background: Clostridioides difficile is the main cause of healthcare-associated diarrhea worldwide. It is proposed that certain C. difficile toxinotypes with distinct pathogenicity locus (PaLoc) variants are associated with disease severity and outcomes. Additionally, few studies have described the common C. difficile toxinotypes, and also little is known about the tcdC variants in Iranian isolates. We characterized the toxinotypes and the tcdC genotypes from a collection of Iranian clinical C. difficile tcdA+B+ isolates with known ribotypes (RTs).Methods: Fifty C. difficile isolates with known RTs and carrying the tcdA and tcdB toxin genes were analyzed. Toxinotyping was carried out based on a PCR-RFLP analysis of a 19.6 kb region encompassing the PaLoc. Genetic diversity of the tcdC gene was determined by the sequencing of the gene.Results: Of the 50 C. difficile isolates investigated, five distinct toxinotypes were recognized. Toxinotypes 0 (33/50, 66%) and V (11/50, 22%) were the most frequently found. C. difficile isolates of the toxinotype 0 mostly belonged to RT 001 (12/33, 36.4%), whereas toxinotype V consisted of RT 126 (9/11, 81.8%). The tcdC sequencing showed six variants (35/50, 70%); tcdC-sc3 (24%), tcdC-A (22%), tcdC-sc9 (18%), tcdC-B (2%), tcdC-sc14 (2%), and tcdC-sc15 (2%). The remaining isolates were wild-types (15/50, 30%) in the tcdC gene.Conclusions: The present study demonstrates that the majority of clinical tcdA+B+ isolates of C. difficile frequently harbor tcdC genetic variants. We also found that the RT 001/ toxinotype 0 and the RT 126/ toxinotype V are the most common types among Iranian isolates. Further studies are needed to investigate the putative association of various tcdC genotypes with CDI severity and its recurrence.

Studies on TLR2 gene variants and their association with milk yield and milk quality traits in Bos indicus (Deoni) cattle

2017 ◽  
Author(s):  
U. T. Mundhe ◽  
D. N. Das ◽  
R. S. Gandhi ◽  
P. Divya
Keyword(s):  
Milk Yield ◽  
Bos Indicus ◽  
Restriction Enzymes ◽  
Milk Quality ◽  
Quality Traits ◽  
Exon 2 ◽  
Allelic Frequencies ◽  
Tlr2 Gene ◽  
Pcr Rflp

Present study molecular characterization of exon 2 of TLR2 gene and its association with milk yield and milk quality traits in 104 Deoni cattle using PCR- RFLP technique was done. Polymorphism was observed through HaeIII, HhaI and EcoRV restriction enzymes in Created Restriction Site (CRS) exon 2-1, CRS exon 2-5 and exon 2-1 by PCR- RFLP, respectively. In CRS exon 2-1 allelic frequencies were observed as 0.793 for A and 0.206 for B alleles and that of genotypic frequencies were 0.58 and 0.41 for genotypes AA and AB. In CRS exon 2-5, two genotypes viz., AC and CC with corresponding allelic frequencies were observed as 0.221 for A and 0.778 for C allele and that of genotypic frequencies observed were 0.44 and 0.55 for AC and CC genotypes respectively. TLR2 exon 2-1 exhibited two alleles G and T with frequencies of 0.134 and 0.865 and their Corresponding genotypic frequencies were 0.009, 0.25 and 0.74for GG, GT and TT genotypes respectively. Higher count of somatic cells (SCC) in TT homozygous and TG heterozygous genotypes, and lower in GG homozygous genotypes were observed in exon 2-1. Strongly significant (P£0.01) effect for least squares means of Test Day milk yield (TDMY) and Somatic Cell Count of CRS exon 2-1 were observed.

scholarly journals Molecular insights into regulation of JAK2 in myeloproliferative neoplasms

Blood ◽  
2015 ◽  
Vol 125 (22) ◽  
pp. 3388-3392 ◽  
Author(s):  
Olli Silvennoinen ◽  
Stevan R. Hubbard
Keyword(s):  
Molecular Characterization ◽  
Human Disease ◽  
Molecular Mechanisms ◽  
Myeloproliferative Neoplasms ◽  
Critical Role ◽  
Hematologic Malignancies ◽  
Janus Kinase ◽  
Constitutive Activation

Abstract The critical role of Janus kinase-2 (JAK2) in regulation of myelopoiesis was established 2 decades ago, but identification of mutations in the pseudokinase domain of JAK2 in myeloproliferative neoplasms (MPNs) and in other hematologic malignancies highlighted the role of JAK2 in human disease. These findings have revolutionized the diagnostics of MPNs and led to development of novel JAK2 therapeutics. However, the molecular mechanisms by which mutations in the pseudokinase domain lead to hyperactivation of JAK2 and clinical disease have been unclear. Here, we describe recent advances in the molecular characterization of the JAK2 pseudokinase domain and how pathogenic mutations lead to constitutive activation of JAK2.

scholarly journals Molecular Analysis of Aquaglyceroporin 1 Gene in Non-Healing Clinical Iso¬lates Obtained from Patients with Cutaneous Leishmaniasis from Central of Iran

2019 ◽  
Author(s):  
Yasaman Alijani ◽  
Saeedeh Sadat Hosseini ◽  
Salman Ahmadian ◽  
Sonia Boughattas ◽  
Gilda Eslami ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cutaneous Leishmaniasis ◽  
Molecular Characterization ◽  
Significance Level ◽  
Detection And Identification ◽  
Pcr Rflp ◽  
Leishmania Spp ◽  
Sensitive Isolate ◽  
The One

Background: Regarding the antimonial-resistant of Leishmania spp., understanding of related mechanism is neces­sary. One of the most important involved molecules is aquaglyceropin1 (AQP1). The aim of this study was molecu­lar analysis of AQP1 gene from antimonial-resistant clinical isolates and its expression. Methods: Overall, 150 patients with cutaneous leishmaniasis referring to the reference laboratories of Yazd and Varzaneh,, located 105km southeast of Isfahan and 240km away from Yazd, were assessed from Jun 2015 to Dec 2017. After sampling, staining was done and evaluated for Leishman by microscope. Samples were collected in RNAlater solution for gene expression analysis in non-healing isolates. DNA extraction was performed from each slide with Leishman body. All patients with L. major isolates detected by ITS1-PCR-RFLP were followed for find­ing the resistant isolates, consequence of molecular characterization of AQP1 using PCR-RFLP. Gene expression of AQP1 from all resistant isolates was assessed in comparison with the one in a sensitive isolate. Statistical analysis was done using SPSS. The significance level was considered ≤0.05. Results: Five isolates were detected as antimonial resistant. Molecular detection and identification were appeared that all were L. major. The molecular characterization of AQP1 showed G562A mutation. Gene expression of AQP1 in resistant isolates showed 1.67 fold higher than the sensitive isolate. Conclusion: We reported a new point mutation of G562A in AQP1 gene involved in molecular mechanism in re­sistant isolates.

scholarly journals An optimized method for the isolation and the molecular characterization of cerebral microvessels in vivo

2018 ◽  
Author(s):  
Yun-Kyoung Lee ◽  
Helen Smith ◽  
Hui Yuan ◽  
Akira Ito ◽  
Teresa Sanchez
Keyword(s):  
Molecular Characterization ◽  
Molecular Mechanisms ◽  
Disease Model ◽  
Enzymatic Digestion ◽  
Cerebral Microvessels ◽  
Rna Integrity ◽  
Bisulfite Treatment ◽  
Mouse Brain Cortex

ABSTRACTThe molecular characterization of cerebral microvessels in experimental disease models has been hindered by the lack of a standardized method to reproducibly isolate intact cerebral microvessels, with consistent cellular compositions, and without the use of enzymatic digestion, which causes undesirable molecular and metabolic changes. Herein, we describe an optimized method for microvessel isolation from mouse brain cortex, which yields microvessel fragments (diameter <50 μm, 89.3% 3-5 μm) with consistent populations of discrete blood-brain barrier components (endothelial cells, pericytes, and astrocyte end feet), retaining high RNA integrity and protein postranslational modifications (e.g. phosphorylation). We demonstrate that this method allows the quantification of changes in gene expression in a disease model (stroke) and the activation of signalling pathways in mice subjected to drug administration. We also describe the isolation of genomic DNA and bisulfite treatment for the assessment of DNA methylation, as well as the optimization of chromatin extraction and shearing from cortical microvessels. Therefore, this protocol will be of great use to improve the understanding of the molecular mechanisms governing cerebrovascular dysfunction, which may help the development of novel therapies for stroke and other neurodegenerative diseases.

scholarly journals Molecular characterization of 'Candidatus Phytoplasma solani' in celery: Case study in Futog

2021 ◽  
Vol 58 (2) ◽  
pp. 66-71
Author(s):  
Tatjana Popović ◽  
Petar Mitrović ◽  
Andrea Kosovac
Keyword(s):  
Molecular Characterization ◽  
Gene Detection ◽  
Convolvulus Arvensis ◽  
Visual Evaluation ◽  
Leaf Yellowing ◽  
Pcr Rflp ◽  
Novi Sad ◽  
Stolbur Phytoplasma

'Candidates Phytoplasma solani', known by its trivial name stolbur phytoplasma, is a plant pathogen infecting numerous crops in Serbia. Celery plants with prominent leaf yellowing and chlorosis, sporadically with tissue necrosis, were observed during August 2020 in Futog, situated in Novi Sad suburbia in Vojvodina. Total of 12 sampled celery plants, 8 symptomatic and 4 asymptomatic ones, were analysed for 'Ca. P. solani' presence. All symptomatic celery plants were infected with stolbur phytoplasma according to the stoll 1 gene detection and therefore were further subjected to multigene molecular characterization on three genes: tuf, stamp and vmpl. Combining molecular tools PCR/RFLP and sequencing reviled two 'Ca. P. solani' multilocus genotypes in celery: tuf-b/Rqg31/V14 and tuf-b/ Rpm35/V14, present in 4 samples each. Obtained results of the strain genotyping are in concordance with previous data on the 'Ca. P. solani' diversity on celery, but supplemented with genotyping of the vmpl gene. Outbreak of stolbur phytoplasma in the assessed locality in Futog is linked to tuf-b epidemiological cycle correlated in Serbia mainly with weed Convolvulus arvensis which was present in the subjected celery plot, and could have been the phytoplasma inoculum source. Visual evaluation of the symptom occurrence suggests on 10-15% of 'Ca. P. solani' affected celery plants scattered throughout the plot corresponding to the pathogen dispersal in crop by cixiid planthoper Hyalesthes obsoletus (Hemiptera: Auchenorrhyncha) associated with C. arvensis, main vector of stolbur phytoplasma in Serbia.

scholarly journals Molecular Characterization of Photoperiodic Flowering in Cultivated Strawberry

HortScience ◽  
2005 ◽  
Vol 40 (4) ◽  
pp. 1105D-1105
Author(s):  
Philip Stewart ◽  
Daniel Sargent ◽  
Thomas Davis ◽  
Kevin Folta
Keyword(s):  
Arabidopsis Thaliana ◽  
Molecular Characterization ◽  
Molecular Mechanisms ◽  
Expression Patterns ◽  
Model Systems ◽  
Day Length ◽  
Photoperiodic Flowering ◽  
Long Day ◽  
Flowering Control

The molecular mechanisms governing photoperiodic flowering has been well defined in the model systems of Arabidopsis thaliana(a facultative long-day plant) and rice (a short-day plant). Photoperiodic flowering control is of great interest to strawberry (Fragaria×ananassa) breeders and growers, and the genetics of photoperiodic flowering have been well studied, indicating that response to day-length is regulated by a small number of genetic loci. Cultivated strawberry is octoploid, so identification of these loci through forward genetic analyses is not practical. Since the componentry of the flowering response is generally conserved between monocots and dicots, we may assume that similar, if not identical, systems are functioning in strawberry as well. The goal of this work is to understand how cultivars likely containing identical photoperiod-sensing components are differentially sensitive to daylength. The expression patterns of genes relevant to the floraltransition were assessed under specific photoperiod conditions to assess similarities and/or differences to the model systems.

scholarly journals Molecular characterization of erythrocyte glycophorin C variants

Blood ◽  
1991 ◽  
Vol 77 (3) ◽  
pp. 644-648
Author(s):  
S Chang ◽  
ME Reid ◽  
J Conboy ◽  
YW Kan ◽  
N Mohandas
Keyword(s):  
Molecular Characterization ◽  
Mechanical Stability ◽  
Exon 2 ◽  
Mrna Sequencing ◽  
Erythrocyte Shape ◽  
Altered Glycosylation ◽  
Exon 1 ◽  
Dna Fragment ◽  
Glycophorin C

Human erythrocyte glycophorin C plays a functionally important role in maintaining erythrocyte shape and regulating membrane mechanical stability. Immunochemical and serologic studies have identified a number of glycophorin C variants that include the Yus, Gerbich, and Webb phenotypes. We report here the molecular characterization of these variants. Amplification of glycophorin C mRNA from the Yus phenotype, using two oligonucleotide primers that span the coding domain, generated a 338-bp fragment compared with a 395-bp fragment generated by amplification of normal glycophorin C mRNA. Sequencing of the mutant 338-bp fragment identified a 57-bp deletion that corresponds to exon 2 of the glycophorin C gene. Similar analysis showed deletion of 84-bp exon 3 in the Gerbich phenotype. In contrast to the generation of shorter than normal DNA fragments from mRNA amplification in the Yus and Gerbich phenotypes, amplification of mRNA from the Webb phenotype generated a normal-sized fragment. Sequencing of this DNA fragment showed an A----G substitution at nucleotide 23 of the coding sequence, resulting in the substitution of asparagine by serine. This modification accounts for the altered glycosylation of glycophorin C seen in this phenotype. These results have enabled us to characterize glycophorin C variants in three different phenotypes that involve deletions of exons 2 and 3 of the glycophorin C gene, as well as a point mutation in exon 1 that results in altered glycosylation of this protein.

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