Identification of point mutation in TLR2 gene and its association with mastitis in water buffalo

2017 ◽  
Author(s):  
Sunita Thakur ◽  
Mahendra Singh ◽  
Gajendra Kumar Aseri ◽  
Archana Verma ◽  
M. R. Vineeth ◽  
...  
Keyword(s):  
Genetic Resistance ◽  
Resistance Mechanisms ◽  
Causative Organism ◽  
Great Loss ◽  
Exon 2 ◽  
Polymorphic Pattern ◽  
Pcr Rflp ◽  
Causative Agents ◽  
Negative Effect ◽  
Potential Strategy

Mastitis is an economically important disease causing great loss to dairy sector owing to its negative effect on production. Its multi factorial etiology makes it difficult to use prophylactic methods for control. Identification of genetic resistance mechanisms and selection for it is a potential strategy for mastitis control. The present study was done in 101 Murrah buffaloes to find the common causative agents of mastitis in buffaloes, to identify variants in exon 2 and 3' UTR of TLR 2 gene and to study their association with intramammary infection. The microbiological examination revealed Staphylococcus aureus to be most common causative organism followed by Streptococcus agalactiae. PCR-RFLP using Sau3AI and HpyCH4V for exon 2 and 3' UTR respectively resulted in polymorphic pattern for exon2, while 3' UTR resulted in monomorphic pattern. PCR RFLP Sau3AI genotypes of TLR2 exon 2 were found to be highly associated with mastitis resistance. AA genotype was found to be least susceptible, while AB genotype was found most frequent among affected buffaloes.

Estudio del polimorfismo y poligenismo de los loci DRB en caballos criollo argentino

2003 ◽  
Author(s):  
◽  
Silvina Díaz
Keyword(s):  
Exon 2 ◽  
Pcr Rflp

El objetivo del trabajo de Tesis Doctoral consistió en estudiar el polimorfismo y el poligenismo de los genes de clase II DRB del Complejo Principal de Histocompatibilidad en Equinos (ELA). La variabilidad genética de las regiones promotoras y de la región de reconocimiento del antígeno (exón 2) se analizó mediante los métodos de PCR-RFLP y PCR-SSCP. Se detectaron tres nuevos alelos del exón 2 definidos por PCR-RFLP, los que se confirmaron por clonado y secuenciación de los productos de amplificación. Además, el número de variantes detectadas en cada animal permitió inferir la presencia de al menos tres copias de genes DRB en los haplotipos equinos analizados. Estos datos se confirmaron a través de análisis filogenético y de segregación. El clonado y secuenciación de la región reguladora (URR) de los genes DRB permitió caracterizar la organización del promotor proximal. Los resultados obtenidos mostraron la presencia en dirección 5´ - 3´ de las cajas conservadas W, X, Y, CCAAT y TATA. Esta estructura es semejante a la reportada en los genes ortólogos de otras especies de mamíferos y evidenciaría que la región analizada corresponde a un gen DRB funcional. Por otra parte, el análisis del polimorfismo del promotor permitió identificar cinco alelos definidos por SSCP. El conjunto de resultados obtenidos mostró que los genes ELA-DRB presentan las principales características descriptas para los genes de Clase II de mamíferos: existencia de copias múltiples, alto grado de polimorfismo, alta tasa de sustituciones no sinónimas en los sitios de reconocimiento del antígeno, y conservación de secuencia y estructura del promotor proximal.

scholarly journals FEATURES OF DIAGNOSIS OF NECROBACTERIOSIS OF COWS BY PCR-RFLP

2020 ◽  
pp. 26-37
Author(s):  
O.D. Biriukova ◽  
T.M. Suprovych ◽  
M.P. Suprovych ◽  
S.V. Laiter-Moskaliuk ◽  
I.O. Chornyi
Keyword(s):  
Molecular Genetic ◽  
Experimental Sample ◽  
Exon 2 ◽  
Blood Samples ◽  
Restriction Fragments ◽  
Black And White ◽  
Pcr Rflp ◽  
Dairy Breed ◽  
Established Technique ◽  
Gene Exon

Molecular genetic markers can detect polymorphism at the DNA level. This feature determines the possibility of their widespread use in genetics and breeding. Alleles of the BoLA-DRB3 gene (exon 2) can act as such markers if a statically significant association between the disease and the allele is established. The presence of such DNA markers in the genotype of animals makes it possible to judge the likelihood of disease in postnatal ontogenesis immediately after the birth of a heifer, based on which we can conclude about the conditions of further use of the animal in the main herd. According to the results of studying the polymorphism of the BoLA-DRB3 gene in cows of the Ukrainian black and white dairy breed resistant and susceptible to necrobacteriosis, four "informative" alleles were revealed. Two of them *03 and *22 are associated with resistance, and the other two - *16 and *23 with susceptibility to necrobacteriosis. The presence of these alleles in the genotype of the animal is determined by testing performed by PCR-RFLP. The method is time consuming, labor intensive and costly. To simplify it, the following technique is proposed. Restriction fragments of alleles *03, *16, *22 and *23 for endocluases RsaI, XhoII and HaeIII have the following DNA patterns: bbb, jbd, mba and nba. Due to the peculiarity of the restriction fragments, which is that endonuclease XhoII reveals in these alleles only one pattern b with length of 284 bp, the process of determining informative alleles can be simplified. Isolation of DNA from blood samples and amplification of a fragment of the BoLA-DRB3.2 gene with a size of 284 bp is carried out according to the established technique. Next, the restriction of the fragment by endonuclease XhoII and sampling having a pattern b. Selected samples are treated with RsaI endonuclease and only those with patterns b, j, m and n remain. The next step is to restrict the selected samples with HaeIII endonuclease and select heifers with bbb (*03) and nba (*23) genotypes. After the first restriction, blood samples without pattern b are eliminated from the experimental sample; after the second – two alleles with patterns RsaI + XhoII jb (*16) and mb (*22) are unambiguously determined, after the third – genotypes bbb and nba, which correspond to alleles *03 and *23. In total, only 75% of blood samples are typed, which reduces the material consumption, time and cost of work to identify heifers genetically susceptible (resistant) to necrobacteriosis.

scholarly journals ATM orchestrates the DNA-damage response to counter toxic non-homologous end-joining at broken replication forks

2018 ◽  
Author(s):  
Gabriel Balmus ◽  
Domenic Pilger ◽  
Julia Coates ◽  
Mukerrem Demir ◽  
Matylda Sczaniecka-Clift ◽  
...  
Keyword(s):  
Genetic Resistance ◽  
Resistance Mechanisms ◽  
Replication Forks ◽  
End Joining ◽  
Strand Breaks ◽  
Dna Damaging Agents ◽  
Genome Wide ◽  
Topoisomerase Poison ◽  
Recombination Repair ◽  
Non Homologous End Joining

SummaryMutations in the ATM tumor suppressor confer hypersensitivity to DNA-damaging agents. To explore genetic resistance mechanisms, we performed genome-wide CRISPR-Cas9 screens in cells treated with the DNA topoisomerase poison topotecan. Thus, we establish that loss of terminal components of the non-homologous end-joining (NHEJ) machinery or the BRCA1-A complex specifically confers topotecan resistance to ATM-deficient cells. We show that hypersensitivity of ATM-mutant cells to topotecan or the poly-(ADP-ribose) polymerase inhibitor olaparib is due to delayed homologous recombination repair at DNA-replication-fork-associated double-strand breaks (DSBs), resulting in toxic NHEJ-mediated chromosome fusions. Accordingly, restoring legitimate repair in ATM-deficient cells, either by preventing NHEJ DNA ligation or by enhancing DSB-resection by BRCA1-A complex inactivation, markedly suppresses this toxicity. Our work suggests opportunities for patient stratification in ATM-deficient cancers and when using ATM inhibitors in the clinic, and identifies additional therapeutic vulnerabilities that might be exploited when such cancers evolve drug resistance.One Sentence SummaryATM counteracts toxic NHEJ at broken replication forks

scholarly journals Circulating Tumour DNA Sequencing Identifies a Genetic Resistance-Gap in Colorectal Cancers with Acquired Resistance to EGFR-Antibodies and Chemotherapy

Cancers ◽  
2020 ◽  
Vol 12 (12) ◽  
pp. 3736
Author(s):  
Franciele H. Knebel ◽  
Louise J. Barber ◽  
Alice Newey ◽  
Dimitrios Kleftogiannis ◽  
Andrew Woolston ◽  
...  
Keyword(s):  
Cancer Cells ◽  
Large Fraction ◽  
Single Agent ◽  
Acquired Resistance ◽  
Genetic Resistance ◽  
Resistance Mutation ◽  
Resistance Mechanisms ◽  
Wild Type ◽  
Primary Resistance ◽  
Clinical Resistance

Epidermal growth factor receptor antibodies (EGFR-Abs) confer a survival benefit in patients with RAS wild-type metastatic colorectal cancer (mCRC), but resistance invariably occurs. Previous data showed that only a minority of cancer cells harboured known genetic resistance drivers when clinical resistance to single-agent EGFR-Abs had evolved, supporting the activity of non-genetic resistance mechanisms. Here, we used error-corrected ctDNA-sequencing (ctDNA-Seq) of 40 cancer genes to identify drivers of resistance and whether a genetic resistance-gap (a lack of detectable genetic resistance mechanisms in a large fraction of the cancer cell population) also occurs in RAS wild-type mCRCs treated with a combination of EGFR-Abs and chemotherapy. We detected one MAP2K1/MEK1 mutation and one ERBB2 amplification in 2/3 patients with primary resistance and KRAS, NRAS, MAP2K1/MEK1 mutations and ERBB2 aberrations in 6/7 patients with acquired resistance. In vitro testing identified MAP2K1/MEK1 P124S as a novel driver of EGFR-Ab resistance. Mutation subclonality analyses confirmed a genetic resistance-gap in mCRCs treated with EGFR-Abs and chemotherapy, with only 13.42% of cancer cells harboring identifiable resistance drivers. Our results support the utility of ctDNA-Seq to guide treatment allocation for patients with resistance and the importance of investigating further non-canonical EGFR-Ab resistance mechanisms, such as microenvironmentally-mediated resistance. The detection of MAP2K1 mutations could inform trials of MEK-inhibitors in these tumours.

Polymorphism Of Exon 2-3 Of Growth Hormone Gene In Surti and Mehsani Goats by PCR-RFLP

2018 ◽  
Vol 8 (5) ◽  
pp. 1
Author(s):  
Jyotishree Bayan ◽  
Vishnu Kharadi ◽  
Umed Ramani ◽  
Mamta Janmeda ◽  
Pawar Vallabh ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Growth Hormone Gene ◽  
Exon 2 ◽  
Pcr Rflp

Integrated Effects of Host Resistance and Insecticide Concentration on Survival of and Turfgrass Damage by the Fall Armyworm (Lepidoptera: Noctuidae)

2004 ◽  
Vol 39 (4) ◽  
pp. 584-597 ◽  
Author(s):  
S. K. Braman ◽  
R. R. Duncan ◽  
W. W. Hanna ◽  
M. C. Engelke
Keyword(s):  
Genetic Resistance ◽  
Residual Activity ◽  
Warm Season ◽  
Fall Armyworm ◽  
Larval Survival ◽  
Resistance Mechanisms ◽  
Lower Survival ◽  
Case Evaluation ◽  
Consumption Rates ◽  
Insecticide Concentration

The residual activity of six concentrations of chlorpyrifos, spinosad, and halofenozide on fall armyworm, Spodoptera frugiperda (J. E. Smith), as mediated by five warm-season turfgrass cultivars expressing varying levels of genetic resistance was evaluated in greenhouse trials. Similarly, varying concentrations of halofenozide were applied to six turfgrass cultivars in the field; mortality of neonate and third-instar fall armyworms was assessed. Reduced rates of chlorpyrifos resulted in lower fall armyworm survival on resistant zoysiagrass cultivars relative to that on bermudagrass or paspalum. In a separate trial when treated with spinosad, survival on the same zoysiagrasses was equal to or greater than that on more susceptible bermuda or paspalum. Reduced rates of halofenozide in another greenhouse trial resulted in lower survival on resistant zoysiagrasses at some concentrations at 7, but not at 14, days exposure compared to more susceptible grasses. In the field, at the full labeled rate of halofenozide, 100% mortality was observed regardless of turfgrass cultivar. Larval survival on the most susceptible turf, “TifEagle”, was higher than that on the remaining turf cultivars at the intermediate rate applied. Larvae exposed to treated turf as third instars displayed a trend toward greater survival at intermediate rates on the two paspalums, “Sea Isle 1” and 561-79, while a trend toward lower survival was observed on “Palisades” and “Cavalier” zoysiagrasses. Factors potentially contributing to the variation in responses observed in the present study include different modes of action of insecticides, host plant resistance mechanisms, differential foliar consumption rates and insecticide dose in relation to body weight. Development of management guidelines for pest management practitioners must address the complexity of potential interactions and may require “case by case” evaluation.

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.

PCR-RFLP Polymorphism of Growth Hormone Gene and Its Association with Growth and Morphometric Traits in Ganjam and Baigani Goats of Odisha

2020 ◽  
Author(s):  
D. K. Karna ◽  
A. Aradhana ◽  
G. D. Nayak ◽  
N. Sahoo ◽  
L. K. Sahoo ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Genetic Variation ◽  
Restriction Enzyme ◽  
Gene Segment ◽  
Growth Hormone Gene ◽  
Morphometric Traits ◽  
Exon 2 ◽  
Genotype Frequencies ◽  
Significant Difference ◽  
Pcr Rflp

Background: Growth Hormone (GH) gene codes for the growth hormone, an anabolic hormone synthesized and secreted by the somatotroph cells of pituitary. Growth hormone influences many processes such as growth, lactation, reproduction and metabolism. Genetic variation in the gene are utilized as markers for selecting animals that are superior in terms growth, production and reproduction traits. Information available on the polymorphism of growth hormone gene of Ganjam and Baigani goats of Odisha is scanty. The current investigation was done to explore the genetic variation of this gene and its association with morphometric traits. Methods: Genetic polymorphism in exon 2 and 3 of Growth hormone gene in Ganjam goat and Baigani goat was explored with sample size of 100 goats for each. The goats belonged to three locations: Khallikote, Rambha, Chattrapur of Ganjam district. The goats were recorded for their body weights, morphometric traits and morphological traits. Genomic DNA was isolated, the target segment comprising exons 2 and 3 was amplified and PCR-RFLP was carried out using Hae III restriction enzyme. Genotypes were scored. Result: PCR of the locus resulted in 422 bp PCR product. PCR-RFLP using Hae III restriction enzyme yielded only two variants in both the populations. The variant A had only one restriction recognition site on the target gene segment yielding two bands with size of 366bp and 56bp whereas the variant B did not have any restriction site with single band of 422bp. Three genotypes AA, AB and BB were found in both the population. In both the population, the gene and genotype frequency were significantly deviated from the Hardy Weinberg Equilibrium frequency. There was a significant difference in the genotype frequencies of growth hormone gene between Ganjam and Baigani goats. The AB genotype had higher mean value for all morphometric traits than AA and BB genotype though the differences were not found to be significant.

scholarly journals Genetic polymorphism and effect on milk production of CSN2 gene in conventional and local cattle breeds in Croatia

Mljekarstvo ◽  
2020 ◽  
Vol 71 (1) ◽  
pp. 3-12
Author(s):  
Ante Ivanković ◽  
Keyword(s):  
Public Interest ◽  
Milk Fat ◽  
Allelic Variation ◽  
The Past ◽  
Cattle Breeds ◽  
Pcr Rflp ◽  
Beta Casein ◽  
Negative Effect ◽  
Central Database

Over the past three decades, the scientific and public interest has been initiated by studies in which the negative effect of milk consumption of beta-casein A1 variant has been observed on consumers’ health. The production of “A2 milk” is one of the ways to economically increase the competitiveness of small and medium-sized dairy farms. Breeders of endangered local breeds are also interested in reaffirmation through production of “A2 milk”. The aim of this study is to determine frequencies of A1 and A2 variants of beta-casein in three commercial and three local breeds of cattle in Croatia, and their relationship with production indicators of commercial breeds in the first three lactations. The genomic DNA was extracted from hair and determination of CSN2 genotypes was conducted by PCR RFLP method. Data for milk yield and chemical composition were provided by the central database. The dominant presence of A2 variant beta-casein in the investigated cattle breeds (0.650-0.758) and the increase in the frequency of A2 beta-casein in the population of Simmental and Istrian cattle were determined. The association of A2A2 and A1A1 genotypes of beta-casein with lactation production and milk fat content in the first and second lactation was observed (p<0.05). The dominance of the A2 allelic variation of beta-casein makes the researched conventional and local breeds suitable for the production of “A2 milk”. The breeding extension of A2 allelic variation of beta-casein within local cattle breeds should be carried out carefully trying not to lose part of the existing genetic variability.

scholarly journals PSII-20 Effects of aerobically exposed corn silage treated with medium chain fatty acids on rumen fermentation in vitro

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 402-403
Author(s):  
Claudio Arzola-Alvarez ◽  
Oscar Ruiz-Barrera ◽  
Yamicela Castillo-Castillo ◽  
Marina Ontiveros ◽  
Mozart Fonseca ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Volatile Fatty Acids ◽  
Corn Silage ◽  
Equal Proportion ◽  
Fermentation Efficiency ◽  
Medium Chain ◽  
Negative Effect ◽  
Pathogenic Microbes ◽  
Potential Strategy

Abstract Medium chain fatty acid (MCFA) treatment of aerobically exposed silage during feed-out is a potential strategy to prevent growth of spoilage and pathogenic microbes. To evaluate the effects of feeding MCFA-treated silage on the rumen fermentability, we incubated 0.02 g of overnight air-exposed corn silage that had been treated individually without or with (0.75% wt/wt) C6, C8, C10 or C12 or with equal proportion mixtures of C6:C8:C10:C12 or C8:C10 with 10 mL freshly collected ruminal fluid. Additions were made while flushing with CO2 and tubes (3 tubes/treatment) were then capped and sealed. After 24 h incubation at 39oC, headspace gases and fluid volatile fatty acids were measure using gas chromatography techniques. Results from a completely randomized analysis of variance revealed higher hydrogen accumulations (P &lt; 0.05) only in incubations containing silage treated with the C8:C10 MCFA mixture when compared to controls, with mean ± SD concentrations (µmol/mL incubation fluid) being 2.19 ± 1.6 versus 0.08 ± 0.04, respectively. Treatment effects were not observed (P &gt; 0.05) on accumulations (µmol/mL) of methane (13.25 ± 2.5), acetate (19.66 ± 5.6), propionate (7.27 ± 2.0), butyrate (12.36 ± 2.7) or stochiometric estimates of amounts of hexose fermented (41.39 ± 9.7), although the acetate:propionate ratios of incubations of silage treated with C8 or the C8:C10 mixture were higher (P &lt; 0.05) than those of untreated controls (3.46 ± 0.7, 3.46 ± 0.5 and 2.35 ± 0.1, respectively). Consequently, correspondingly estimates of fermentation efficiency, calculated based caloric energy values of amounts of acetate, propionate and butyrate relative to glucose, were slightly lower (P &lt; 0.05) for the incubations of silage treated with C8 and the C8:C10 mix compared to controls (74.36 ± 1.1, 74.82 ± 0.2 and 76.55 ± 0.3 %, respectively). Results reveal little negative effect of MCFA treatment on ruminal fermentability of silage.

Sign in / Sign up

Export Citation Format

Share Document