scholarly journals Nucleotide Sequencing and SNP Detection of Toll-Like Receptor-4 Gene in Murrah Buffalo (Bubalus bubalis)

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
M. Mitra ◽  
S. Taraphder ◽  
G. S. Sonawane ◽  
A. Verma
Keyword(s):  
Rflp Analysis ◽  
Ovis Aries ◽  
Capra Hircus ◽  
Nucleotide Sequencing ◽  
Toll Like Receptor ◽  
Snp Detection ◽  
Toll Like Receptor 4 ◽  
Exon 2 ◽  
Murrah Buffalo ◽  
Pcr Rflp

Toll-like receptor-4 (TLR-4) has an important pattern recognition receptor that recognizes endotoxins associated with gram negative bacterial infections. The present investigation was carried out to study nucleotide sequencing and SNP detection by PCR-RFLP analysis of the TLR-4 gene in Murrah buffalo. Genomic DNA was isolated from 102 lactating Murrah buffalo from NDRI herd. The amplified PCR fragments of TLR-4 comprised of exon 1, exon 2, exon 3.1, and exon 3.2 were examined to RFLP. PCR products were obtained with sizes of 165, 300, 478, and 409 bp. TLR-4 gene of investigated Murrah buffaloes was highly polymorphic with AA, AB, and BB genotypes as revealed by PCR-RFLP analysis using Dra I, Hae III, and Hinf I REs. Nucleotide sequencing of the amplified fragment of TLR-4 gene of Murrah buffalo was done. Twelve SNPs were identified. Six SNPs were nonsynonymous resulting in change in amino acids. Murrah is an indigenous Buffalo breed and the presence of the nonsynonymous SNP is indicative of its unique genomic architecture. Sequence alignment and homology across species using BLAST analysis revealed 97%, 97%, 99%, 98%, and 80% sequence homology with Bos taurus, Bos indicus, Ovis aries, Capra hircus, and Homo sapiens, respectively.

scholarly journals Toll-like receptor-4 gene variations in Egyptian children with familial Mediterranean fever

2021 ◽  
Vol 48 (1) ◽  
Author(s):  
Yomna Farag ◽  
Samia Salah ◽  
Hanan Tawfik ◽  
Mai Hamed ◽  
Huda Marzouk
Keyword(s):  
Familial Mediterranean Fever ◽  
Disease Severity ◽  
Gene Mutation ◽  
Rflp Analysis ◽  
Gene Variants ◽  
Gene Variant ◽  
Toll Like Receptor ◽  
Toll Like Receptor 4 ◽  
Egyptian Children ◽  
Mediterranean Fever

Abstract Background Familial Mediterranean fever (FMF) is an autosomal recessive disorder affecting people in the region of the Mediterranean Sea. It is usually associated with mutation in Mediterranean fever (MEFV) gene that encodes the pyrin protein, which affects the innate inflammatory response. Toll-like receptors (TLR) are a family of pattern recognition receptors that recognize pathogenic microbes and activate antimicrobial defense mechanisms. Toll-like receptor 4 (TLR-4) is concerned with recognition of gram-negative organisms. There is growing clinical evidence suggesting a role for expression of TLRs in the immune pathogenesis of FMF. Thus, the aim of the current study was to evaluate the presence of TLR-4 (p.Asp299Gly) and TLR-4 (p.Thr399Ile) gene variants in association with Egyptian children having FMF, furthermore, its effect on disease course and severity. Results Seventy Egyptian children diagnosed as having FMF, together with 50 age and gender-matched controls were enrolled in the study. The TLR-4 (p.Asp299Gly) and (Thr399Ile) gene variants were determined by PCR-RFLP analysis for all studied patients and controls. TLR-4 p.Asp299Gly gene variant was detected in 1 (1.4%) of the patients and p.Thr399Ile gene variant was detected in 2 (2%). None of the controls had any of the two tested gene variants. All found variations were heterozygous. We could not find a statistically significant association with disease severity in cases with or without TLR-4 gene variants (P = 0.568). Patients with M694V gene mutation showed a higher disease severity (P = 0.035). Conclusion TLR-4 (p.Asp299Gly) and (p.Thr399Ile) gene variants were not found to have a link with the occurrence, the clinical picture of FMF, its severity, and response to colchicine treatment in Egyptian children. M694V gene mutation seems to be associated with higher disease severity. Further larger studies are needed to verify these results.

scholarly journals TACI και σήψη σε βαρέως πάσχοντες ασθενείς στη ΜΕΘ

2014 ◽  
Author(s):  
Μαρία Κομπότη
Keyword(s):  
Sofa Score ◽  
Hazard Ratio ◽  
Apache Ii ◽  
Apache Ii Score ◽  
Toll Like Receptor ◽  
Wild Type ◽  
Odds Ratios ◽  
Toll Like Receptor 4 ◽  
Pcr Rflp

Σκοπός: ο προσδιορισμός των γενετικών πολυμορφισμών των μορίων TACI, BAFF και TLR4 και η διερεύνηση πιθανής συσχέτισής τους με την εμφάνιση σήψης και τη θνητότητα σε ασθενείς που εισάγονται στη ΜΕΘ. Ασθενείς-Μέθοδος: Προοπτική μελέτη παρακολούθησης όλων των ασθενών που εισήχθησαν στη Μονάδα Εντατικής Θεραπείας μέχρι την έκβαση (έξοδος, θάνατος). Για τη φυσική ανοσία μελετήθηκαν δύο πολυμορφισμοί του toll-like receptor 4 (ΤLR4-D299G και TLR4-T399I) και ένας πολυμορφισμός του C2 που προκαλεί ανεπάρκεια συμπληρώματος (C2del28bp). Για την επίκτητη ανοσία μελετήθηκαν δύο πολυμορφισμοί του BAFF-R (BAFF-R-H159Y και BAFF-R-P21R) και ένας του TACI (TACI-C104R). Η ανίχνευση των πολυμορφισμών έγινε με αλυσιδωτή αντίδραση πολυμεράσης και ανάλυση με ενδονουκλεάσες περιορισμού (PCR-RFLP). Αποτελέσματα: Στη μελέτη συμπεριελήφθησαν 215 ασθενείς (148 άνδρες και 67 γυναίκες). Η ηλικία (μέσος ± SD) ήταν 54,1±19,7 έτη, APACHE II score εισαγωγής στη ΜΕΘ 22,0±6,0 και SOFA score εισαγωγής στη ΜΕΘ 9,7±3,5. Σήψη στη ΜΕΘ βρέθηκε σε 108 ασθενείς (50,2%) και η θνητότητα στη ΜΕΘ ήταν 20,5% [95% διάστημα αξιοπιστίας 15,0–25,9]. Οι SNPs του TLR4 βρέθηκαν σε συζευγμένη κατάσταση. Οι φορείς κάποιου TLR4 SNP και οι φορείς του BAFFR-P21R συσχετίστηκαν ανεξάρτητα με χαμηλότερη πιθανότητα σήψης στη ΜΕΘ συγκριτικά με τους wild-type ομοζυγώτες [διορθωμένα odds ratios 0,32, 95%CI 0,12–0,86, p=0,024 για τον TLR4-T399I, 0,34, 95%CI 0,13–0,94, p=0,037 για τον TLR4-T399I and 0,44, 95%CI 0,20–0,97, p=0,044 για τον BAFFR-P21R]. Από την ανάλυση υποομάδων, φάνηκε ότι η συσχέτιση αυτή αφορούσε στους παθολογικούς ασθενείς και οριακά στους τραυματίες, ενώ στους χειρουργικούς ασθενείς δεν παρατηρήθηκε συσχέτιση. Η φορεία του TACI-C104R SNP είχε καλή προβλεπτική αξία μετά από διόρθωση ως προς συγχυτικούς παράγοντες [hazard ratio 5,01 (1,14–22,03, p=0,033). Ο BAFFR-H159Y SNP δεν συσχετίστηκε με την εμφάνιση σήψης και τη θνητότητα, ενώ ο πολυμορφισμός C2del28bp δεν ανευρέθηκε στο δείγμα της μελέτης.Συμπεράσματα: Η μελέτη μας έδειξε ότι σε βαρέως πάσχοντες ασθενείς ΜΕΘ, οι πολυμορφισμοί TLR4-D299G (rs4986790), TLR4-T399I (rs4986791) και TNFRSF13C/BAFFR-P21R (rs77874543) ασκούν προστατευτική επίδραση αναφορικά με την εμφάνιση σήψης, ενώ ο πολυμορφισμός TNFRSF13B/TACI-C104R (rs34557412) συσχετίζεται σημαντικά με αύξηση της θνητότητας στη ΜΕΘ. Ο πολυμορφισμός TNFRSF13C/BAFFR-H159Y (rs61756766) δεν συσχετίστηκε με την εμφάνιση σήψης ή τη θνητότητα στη ΜΕΘ. Διαπιστώθηκε ανισορροπία σύνδεσης για τους πολυμορφισμούς TLR4-D299G (rs4986790) και TLR4-T399I (rs4986791) και για τους πολυμορφισμούς TNFRSF13C/BAFFR-P21R (rs77874543) και TNFRSF13C/BAFFR-H159Y (rs61756766). Ο πολυμορφισμός C2-c.841_849+19del28 (rs9332736) δεν ανιχνεύτηκε στο δείγμα των ασθενών της μελέτης.

PCR-RFLP Authentication of Meats from Red Deer (Cervus elaphus), Fallow Deer (Dama dama),Roe Deer (Capreolus capreolus), Cattle (Bos taurus), Sheep (Ovis aries), and Goat (Capra hircus)

2006 ◽  
Vol 54 (4) ◽  
pp. 1144-1150 ◽  
Author(s):  
Violeta Fajardo ◽  
Isabel González ◽  
Inés López-Calleja ◽  
Irene Martín ◽  
Pablo E. Hernández ◽  
...  
Keyword(s):  
Cervus Elaphus ◽  
Red Deer ◽  
Roe Deer ◽  
Bos Taurus ◽  
Capreolus Capreolus ◽  
Fallow Deer ◽  
Ovis Aries ◽  
Capra Hircus ◽  
Dama Dama ◽  
Pcr Rflp

scholarly journals Fatal systemic toxoplasmosis in a 3-month-old young tibetan goat (Capra hircus)

2020 ◽  
Vol 16 (1) ◽  
Author(s):  
Silvia Pavone ◽  
Silvia Crotti ◽  
Deborah Cruciani ◽  
Nicoletta D’Avino ◽  
Jacopo Zema ◽  
...  
Keyword(s):  
Histopathological Examination ◽  
Fatal Case ◽  
Fatal Outcome ◽  
Rflp Analysis ◽  
Capra Hircus ◽  
Parasitic Infections ◽  
Type Ii ◽  
Intermediate Hosts ◽  
Pcr Rflp ◽  
Generalized Lymphadenopathy

Abstract Background Toxoplasmosis is one of the most common parasitic infections in both humans and animals. It is a frequent cause of abortion and stillbirth in intermediate hosts, especially sheep and goats but rarely causes fatal clinical form in adult animals. Case presentation In contrast, the study reports an unusual fatal case of toxoplasmosis in a young goat naturally infected with type II strain of Toxoplasma gondii. A three-month-old female goat was presented with dyspnea and died few days later. Grossly, lungs were firm, edematous and mottled with disseminated whitish areas. Generalized lymphadenopathy was found. The histopathological examination showed necrotic interstitial bronchopneumonia and necrotizing lymphadenitis with intralesional free and clustered within macrophages tachyzoites of T. gondii. DNA extracted from lungs and lymph nodes was positive for T. gondii by a fast qPCR. PCR-RFLP analysis and sequencing of GRA6 gene showed that the isolated strains belonged to type II genotype. Conclusions This is an unusual report of acute systemic toxoplasmosis caused by the type II strain of T. gondii with a fatal outcome in a young goat.

scholarly journals Genetic heterogeneity among parapoxviruses isolated from sheep, cattle and Japanese serows (Capricornis crispus)

2001 ◽  
Vol 82 (5) ◽  
pp. 1215-1220 ◽  
Author(s):  
Yasuo Inoshima ◽  
Kenji Murakami ◽  
Takashi Yokoyama ◽  
Hiroshi Sentsui
Keyword(s):  
Fragment Length Polymorphism ◽  
Genetic Characterization ◽  
Rflp Analysis ◽  
Nucleotide Sequencing ◽  
Envelope Gene ◽  
Viral Dna ◽  
Pcr Rflp ◽  
Bovine Papular Stomatitis Virus ◽  
Restriction Fragment Length

Standard strains of four parapoxviruses and seven unclassified Japanese strains isolated from sheep, cattle and wild Japanese serows (Capricornis crispus) were compared molecularly. Restriction fragment length polymorphism (RFLP) analysis of viral DNA, indirect immunofluorescence assays using monoclonal antibodies, partial nucleotide sequencing of the envelope gene, phylogenetic analysis and PCR–RFLP were carried out. These analyses revealed that the parapoxviruses were divided into four groups and the region sequenced in this study was highly conserved within each group. Each of the Japanese isolates was classified into one of these groups. These findings also indicated that parapoxvirus infections among wild Japanese serows seem to be caused by at least two different parapoxviruses, bovine papular stomatitis virus and orf virus. The methods presented here are useful for genetic characterization and classification of parapoxviruses.

Two Novel Splice Variants of the Ovis aries Toll-like Receptor 4

2011 ◽  
Vol 6 (2) ◽  
pp. 155-165 ◽  
Author(s):  
Bao-Lu Zhang ◽  
Jin-ling Du ◽  
Hong-Bing Han ◽  
Zheng-xing Lian ◽  
Ning Li
Keyword(s):  
Splice Variants ◽  
Ovis Aries ◽  
Toll Like Receptor ◽  
Toll Like Receptor 4

Detection of polymorphism of Calgranulin A gene in Indian Murrah Buffalo

2015 ◽  
Author(s):  
Sourabh Sulabh ◽  
Archana Verma ◽  
I. D. Gupta ◽  
S. Rajesh Kumar
Keyword(s):  
Immune System ◽  
Genetic Variants ◽  
Genomic Dna ◽  
Annealing Temperature ◽  
Rflp Analysis ◽  
Clinical Mastitis ◽  
Murrah Buffalo ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Calgranulin A

Calgranulin A (S100A8) gene is one of the important candidate genes, which affects the host disease resistance by enhancing the immune system. Present study was undertaken with the objectives to identify polymorphism in Calgranulin A gene and to associate identified genetic variants with the incidence of clinical mastitis in Murrah buffalo. Genomic DNA was isolated from 100 randomly selected lactating Murrah. Two sets of primers were designed to amplify targeted regions of the gene. PCR products were obtained at annealing temperature of 63.8oC and were of 449 and 489 bp for respective primer set. PCR-RFLP analysis was carried out using HinfI for contig I and AluI and MboII restriction endonucleases for contig II. Both the contigs revealed monomorphism with frequency of the only prevailing A allele as 1.00. It was not feasible to analyze association with the incidence of mastitis, as no genetic variants were observed in the animals studied.

scholarly journals Comparative analysis of IGFBP-3 gene sequence in Egyptian sheep, cattle, and buffalo

2019 ◽  
Vol 12 (1) ◽  
Author(s):  
Ahmed. A. Saleh ◽  
Amr M. A. Rashad ◽  
Nada. N. A. M. Hassanine ◽  
Mahmoud A. Sharaby ◽  
Yongju Zhao
Keyword(s):  
Comparative Analysis ◽  
Gene Sequence ◽  
Restriction Enzymes ◽  
Restriction Pattern ◽  
Nucleotide Sequencing ◽  
Sequencing Analysis ◽  
Exon 2 ◽  
Pcr Rflp ◽  
Digestion Profile ◽  
Igfbp 3

Abstract Objective A total of 205 animals from four Egyptian livestock species; cattle (n = 18), buffaloes (n = 12), sheep (n = 150) and goats (n = 25) were used in this study to detect polymorphism and perform comparative analysis for IGFBP-3 gene using DNA sequencing and (PCR–RFLP). Results The amplified fragments were found to be of length 654 bp in sheep, 651 bp in cattle and 655 bp in buffalo. For Falahy goats, PCR was performed to amplify a 316 bp fragment from exon 2 of the IGFBP-3 gene. The digestion of 654 bp with HaeIII restriction enzyme yielded a single restriction pattern for goats, while for cattle, 3 genotypes were identified; (AA), (AB), and (BB). Moreover, for buffalo one genotype (AA) only was found with HaeIII and TaqI restriction enzymes, separately. Also, the digestion profile for goats with HaeIII revealed one pattern only. Nucleotide sequencing of the amplified fragments of IGFBP-3 gene in sheep, cattle, buffalo, and goat was submitted to the NCBI GenBank (Accession no. MG738671.1, MG738673.1, MG738674.1, and MG738672.1, respectively). The nucleotide sequencing analysis indicated similarity percentages in IGFBP-3 gene fragments of 88.54, 89.63 and 95.06% between “sheep and cattle”, “sheep and buffalo”, and “cattle and buffalo”, respectively.

scholarly journals Identification of BMP15 Exon 2 for fecundity traits by PCR-RFLP and nucleotide sequencies in Kejobong goat

2017 ◽  
Vol 42 (4) ◽  
pp. 220 ◽  
Author(s):  
A. Febriana ◽  
S. Sutopo ◽  
E. Kurnianto
Keyword(s):  
Insertion Site ◽  
Pcr Amplification ◽  
Nucleotide Sequencing ◽  
Specific Marker ◽  
Exon 2 ◽  
High Productivity ◽  
Morphogenetic Protein ◽  
Pcr Rflp ◽  
Sequencing Technique ◽  
Gene Exon

Kejobong goat is known as prolific and high productivity goat breed in Indonesia. PCR-RFLP and sequencing technique was established in the present study to accomplish the polymorphisms of Bone Morphogenetic Protein 15 (BMP15) gene exon 2 on Kejobong goat does. The blood samples was collected from 48 Kejobong does which were selected based on their litter size. The size of PCR amplification of BMP15 gene exon 2 was 837 bp. The product of PCR-RFLP technique digested by HinfI enzyme showed that the samples were monomorphic. Authentication result using nucleotide sequencing found 4 substitution (A391G, C464G, T828C and C830G), 1 alignment gap (site 817) and 1 insertion nucleotide (site 822). This mutations caused 6 haplotypes formatted. The mutants of BMP15 exon 2 on Kejobong goats indicated that this breed had their own mutation controling the prolific trait. The phylogenetic tree build on the sequences of BMP15 gene exon 2 of Kejobong goats was grouped into 3 clusters. The alignment gap indicated to be the specific marker for the prolific trait (duplet) in Kejobong goat. The particular insertion site could be the recognition site of Kejobong goat based on BMP15 exon 2.

scholarly journals Comparitive Genome Sequence Analysis of Bovine Lymphocyte Antigen BoLA DRB3.2 Alleles in Deoni and Ongole (Bos indicus) Cattle Breeds of India

2021 ◽  
Author(s):  
R. Saravanan ◽  
N. Murali ◽  
A.K. Thiruvenkadan ◽  
D.N. Das
Keyword(s):  
Sequence Analysis ◽  
Direct Sequencing ◽  
Major Gene ◽  
Rflp Analysis ◽  
Mobility Shift ◽  
Exon 2 ◽  
Lymphocyte Antigen ◽  
Pcr Rflp ◽  
Bovine Lymphocyte ◽  
Indian Cattle

Background: India, a major livestock region of the Asian countries is rich in animal genetic resources having special qualities of hardy nature, resistance to many diseases and adopted to adverse climatic conditions. The cattle MHC, Bovine Lymphocyte Antigen DRB3 (BoLA-DRB3) is considered to be a major gene linked with disease resistance traits of Indian cattle. Methods: The present study was carried out to sequence the BoLA-DRB3.2 alleles in Deoni and Ongole breeds of Indian cattle. PCR RFLP analysis of the BoLA-DRB3.2 alleles in Deoni (n=51) and Ongole (n=60) cattle using three different restriction enzymes RsaI, BstYI and HaeIII to find out the possible restriction pattern. Based on the combined allelic patterns, each sample was further analyzed by PCR- SBT technique to detect the SNP variations present in BoLA-DRB3.2 alleles.Result: The PCR RFLP analysis revealed that the highest frequent alleles are *6 (0.216) and *15 (0.225) in Deoni and Ongole breeds of cattle, respectively. The second-highest frequency was observed for BoLA alleles *11 and *6 which were present at a frequency of 0.167 and 0.200 in Deoni and Ongole breeds of cattle, respectively. To get the complete picture of polymorphic pattern of BoLA-DRB3.2 allele direct sequencing was carried out for each plymorphic pattern. The interesting feature noticed in the Ongole breed was that at position 91 and 133 of the sequence, it had both A and G nucleotides in contrast to Bos taurus breed, which had only TT nucleotides. The sequence analysis of BoLA-DRB3 exon 2 between two breeds revealed that there are numerous variations in exon 2, whatever variation, that lead to different mobility shift and band pattern in gels. Deoni and Ongole breeds of cattle had similar variations at positions 94, 134, 211, 235 and 258noticed due to the unique nature of native breeds.

Sign in / Sign up

Export Citation Format

Share Document