scholarly journals PSIV-2 Investigating candidate scur genes in Bos taurus breeds

2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 226-227 ◽  
Author(s):  
Crystal Ketel ◽  
Mika Asai-Coakwell
Keyword(s):  
Candidate Genes ◽  
Gel Electrophoresis ◽  
Bos Taurus ◽  
Pcr Amplification ◽  
Genetic Mutation ◽  
Mapping Data ◽  
Future Studies ◽  
Feedlot Steers ◽  
Pcr Rflp ◽  
Beef Producers

Abstract Scurs (loose horns) are inherited in a sex-influenced manner and are believed to appear in cattle when the animal is heterozygous (Pp) for the polled mutation. They are unwanted by beef producers, but are difficult to eradicate because scurs are epistatic to the polled mutation. With the development of a test for the 202 bp indel on BTA1 resulting in the polled phenotype in Celtic breeds, horned (pp) and polled (PP/Pp) animals can be genotyped at this locus. The aims of this study were: 1) to confirm the polled genotype in scurred families from a Canadian beef research herd (SCBRH), scurred cattle families from producers (SCFP), and polled and scurred feedlot steers using the Celtic poll test (PC), and 2) to identify new candidate genes between the recombinant genes of the postulated scur loci on BTA19. Through PCR amplification, the polled/horned genotype was confirmed in the SCBRH, SPCF, and 153 phenotyped feedlot steers with gel electrophoresis. One family from the SPCF, 26 scurred and 10 polled feedlot steers were genotyped as horned. Removing the SPCF horned animals from the scur loci mapping data changed the recombinant markers and created a new boundary, resulting in examination of five new candidate genes (CTDNEP1, FGF11, SOX15, SHBG, DHRS7C) based on function and position. To identify SNPs segregating with scurs, 16 animals were chosen from the PC genotyped feedlot steers, 8 Pp scurred steers and 8 Pp polled steers. Two SNP’s found in CTDNEP1 and DHRS7C were examined in the SCBRH with PCR-RFLP using BseRI and AciI, respectively, but did not segregate with scurs. In conclusion, careful phenotyping and genotyping for the polled/horned status of an animal should be confirmed for future studies to determine the genetic mutation resulting in scurs.

PCR-based molecular discrimination betweenMaritrema eroliaeandProbolocoryphe uca(Digenea: Microphallidae) in Kuwait Bay

2013 ◽  
Vol 88 (2) ◽  
pp. 177-182
Author(s):  
W.Y. Al-Kandari ◽  
S.A. Al-Bustan ◽  
M. Alnaqeeb ◽  
A.M. Isaac
Keyword(s):  
Fragment Length Polymorphism ◽  
Pcr Amplification ◽  
Direct Pcr ◽  
Specific Primers ◽  
Kuwait Bay ◽  
Future Studies ◽  
Larval Stages ◽  
Marine Snails ◽  
Pcr Rflp ◽  
Species Specific

AbstractMicrophallid trematodes are common parasites in marine snails and crustacean hosts at Kuwait Bay. The larval stages of two microphallids,Maritrema eroliaeandProbolocoryphe uca, are difficult to differentiate morphologically. In this study, two PCR-based techniques were established for quick and accurate discrimination between the larval stages of the two microphallid species, employing restriction fragment length polymorphism (PCR-RFLP) and species-specific primers. Both techniques utilized nucleotide differences in the second internal transcribed region (ITS2) of the ribosomal DNA (rDNA) in the two species. For the PCR-RFLP technique, restriction enzymeAvaII was selected and it generated different restriction profiles among the two microphallids. In addition, species-specific primers were prepared for each microphallid species that amplified distinctive fragments. Both techniques showed that the larval stages of the two microphallid species can be identified accurately. However, direct PCR amplification using species-specific primers was more advantageous than the PCR-RFLP technique since it allowed rapid and specific discrimination between the two species. This technique provides a useful tool that can be used in future studies for the study of the distribution of microphallid species and their definitive hosts at different localities of Kuwait Bay.

Simple and Rapid Detection of Factor V Leiden by Allele-specific PCR Amplification

1996 ◽  
Vol 75 (05) ◽  
pp. 757-759 ◽  
Author(s):  
Rainer Blasczyk ◽  
Markus Ritter ◽  
Christian Thiede ◽  
Jenny Wehling ◽  
Günter Hintz ◽  
...  
Keyword(s):  
Direct Sequencing ◽  
Activated Protein C ◽  
Factor V Leiden ◽  
Pcr Amplification ◽  
Factor V ◽  
Specific Pcr ◽  
Pcr Rflp ◽  
Allele Specific ◽  
Specific Amplification ◽  
Allele Specific Pcr

SummaryResistance to activated protein C is the most common hereditary cause for thrombosis and significantly linked to factor V Leiden. In this study, primers were designed to identify the factor V mutation by allele-specific PCR amplification. 126 patients with thromboembolic events were analysed using this technique, PCR-RFLP and direct sequencing. The concordance between these techniques was 100%. In 27 patients a heterozygous factor VGln506 mutation was detected, whereas one patient with recurrent thromboembolism was homozygous for the point mutation. Due to its time- and cost-saving features allele-specific amplification should be considered for screening of factor VGln506.

Expression of candidate genes for residual feed intake in tropically adapted Bos taurus and Bos indicus bulls under thermoneutral and heat stress environmental conditions

2021 ◽  
pp. 102998
Author(s):  
Bianca Vilela Pires ◽  
Nedenia Bonvino Stafuzza ◽  
Luara Afonso de Freitas ◽  
Maria Eugênia Zerlotti Mercadante ◽  
Ester Silveira Ramos ◽  
...  
Keyword(s):  
Heat Stress ◽  
Candidate Genes ◽  
Feed Intake ◽  
Environmental Conditions ◽  
Bos Taurus ◽  
Bos Indicus ◽  
Residual Feed Intake

scholarly journals PCR-based genotyping assays to detect germline APC variant associated with hereditary gastrointestinal polyposis in Jack Russell terriers

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Kyoko Yoshizaki ◽  
Akihiro Hirata ◽  
Hiroyuki Matsushita ◽  
Naohito Nishii ◽  
Mifumi Kawabe ◽  
...  
Keyword(s):  
Mutant Allele ◽  
False Negative ◽  
Disease Onset ◽  
Pcr Amplification ◽  
Buccal Swab ◽  
Wild Type ◽  
Gastrointestinal Polyposis ◽  
Pcr Rflp ◽  
Formalin Fixed Paraffin Embedded ◽  
Rflp Assay

Abstract Background The prevalence of gastrointestinal (GI) neoplastic polyps in Jack Russell terriers (JRTs) has increased in Japan since the late 2000s. Recently, we demonstrated that JRTs with GI polyps harbor identical germline variant in the APC gene (c.[462_463delinsTT]) in the heterozygous state. Thus, this disease is an autosomal dominant hereditary disorder. Although the affected JRTs have distinct features, such as the development of multiple GI polyps and an early age of disease onset, genetic testing is indispensable for a definitive diagnosis. Here, polymerase chain reaction (PCR)-based assays capable of detecting germline APC variant were designed and validated using synthetic wild-type and mutant DNAs and genomic DNAs from carrier and non-carrier dogs. Result First, the PCR-restriction fragment length polymorphism (PCR-RFLP) assay was developed by taking advantage of the germline APC variant creating a new restriction site for MseI. In the PCR-RFLP assay, the 156-bp region containing the variant site was amplified by PCR and subsequently digested with MseI, yielding diagnostic 51 and 58 bp fragments from the mutant allele and allowing determination of the APC genotypes. It was possible to determine the genotypes using genomic DNA extracted from the peripheral blood, buccal swab, or formalin-fixed paraffin-embedded tissue. Next, a TaqMan duplex real-time PCR assay was developed, where a 78-bp region flanking the variant was amplified in the presence of wild-type allele- and mutant allele-specific fluorescent probes. Using blood-derived DNA, altogether 40 cycles of PCR amplification determined the APC genotypes of all examined samples by measuring the fluorescence intensities. Importantly, false-positive and false-negative errors were never detected in both assays. Conclusion In this study, we developed highly reliable genetic tests for hereditary GI polyposis in JRTs, providing accurate assessment of the presence of the causative germline APC variant. The genotyping assays could contribute to the diagnosis and prevention of hereditary GI polyposis in dogs.

Identification of novel candidate genes for age at first calving in Nellore cows using a SNP chip specifically developed for Bos taurus indicus cattle

2021 ◽  
Author(s):  
Miguel Angel Carabantes Dubom ◽  
Victor Breno Pedrosa ◽  
Fabieli Loise Braga Feitosa ◽  
Raphael Bermal Costa ◽  
Gregório Miguel Ferreira de Camargo ◽  
...  
Keyword(s):  
Candidate Genes ◽  
Bos Taurus ◽  
Age At First Calving ◽  
Snp Chip ◽  
Bos Taurus Indicus

scholarly journals Identification of restriction enzyme in the FSHR gene of indonesian local cattle

2021 ◽  
Vol 888 (1) ◽  
pp. 012024
Author(s):  
P W Prihandini ◽  
A Primasari ◽  
M Luthfi ◽  
D Pamungkas ◽  
A P Z N L Sari ◽  
...  
Keyword(s):  
Restriction Enzyme ◽  
Restriction Site ◽  
Follicle Stimulating Hormone ◽  
Restriction Enzymes ◽  
Future Studies ◽  
Fshr Gene ◽  
Pcr Rflp ◽  
Mapping Analysis ◽  
Pcr Products ◽  
Enzyme Mapping

Abstract The restriction enzyme is important for genotyping using the PCR-RFLP technique. Therefore, this study aims to identify the restriction enzyme mapping in the partial sequence of the follicle-stimulating hormone receptor (FSHR) gene in Indonesian local cattle. A total of 29 samples sized 306 bp, were aligned with Genbank sequence acc no. NC_032660, resulting three polymorphic sites, namely g.193G>C, g.227T>C, and g.275A>C. Furthermore, the restriction mapping analysis using the NEBcutter program V2.0 showed that no enzyme recognized the SNP g.275A>C, while the SNP g.193G>C and g.227T>C were identified by the AluI and MscI enzymes, respectively. The AluI enzyme cuts at two positions (193 bp and 243 bp) in the G allele sample producing three fragments namely 50 bp, 63 bp, and 193 bp, meanwhile, in the C allele, the AluI cuts only in position 243 bp, hence, the fragment products are 63 bp and 243 bp. In contrast, the MscI enzyme was only recognized in the T allele, producing fragments sized 77 bp and 229 bp but failed to identify the restriction site along with the PCR products in the C allele. Based on the results, the SNPs (g.193G>C and g.227T>C) and restriction enzymes (AluI and MscI) are applicable for genotyping local Indonesian cattle using the PCR-RFLP technique in future studies.

scholarly journals Genetic polymorphism of candidate genes in pig meat production

2011 ◽  
pp. 37-40
Author(s):  
Zuzana Lieskovská ◽  
Anton Kováčik ◽  
Anna Trakovická
Keyword(s):  
Genetic Variation ◽  
Genetic Polymorphism ◽  
Candidate Genes ◽  
Fat Content ◽  
Meat Production ◽  
Lepr Gene ◽  
Pcr Rflp ◽  
Pig Meat

H-FABP, LEPR and MC5R genes were suggested as candidate genes for fat content in pig meat. The aim of this study was to detect genetic variation in the porcine H-FABP, LEPR and MC5R genes by PCR-RFLP method in a group of pigs. Genotyping of pigs was done by PCRRFLP methods. We identified three genotypes in the set of pigs, HH (0.504), Hh (0.412) and hh (0.084) for H-FABP (HinfI). Allele H showed higher frequency than allele h (0.710 vs. 0.290). Three genotypes were identified for the H-FABP (HaeIII) gene (DD - 0.194, Dd - 0.494, dd - 0.312). The allele D (0.441) showed slightly lower frequency than allele d (0.559). All three genotypes were identified for LEPR (HpaII) in the group of pigs (AA – 0.137, AB - 0.314, BB – 0.549). Higher frequency of LEPR gene was confirmed for allele B (0.706), as compared with allele A (0.294). We identified two genotypes for MC5R (BsaHI) in the group of pigs (AA - 0.348 and AG - 0.652), genotype GG was not found. As conforms with genotype structure, we recognize a higher frequency of allele A (0.674) as compared with allele G (0.326). 

Coordinate amplification of metallothionein I and II genes in cadmium-resistant Chinese hamster cells: implications for mechanisms regulating metallothionein gene expression

1985 ◽  
Vol 5 (2) ◽  
pp. 320-329
Author(s):  
B D Crawford ◽  
M D Enger ◽  
B B Griffith ◽  
J K Griffith ◽  
J L Hanners ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Chinese Hamster ◽  
Chinese Hamster Cell ◽  
Chinese Hamster Cells ◽  
Future Studies ◽  
Noncoding Regions ◽  
Genes Encoding ◽  
Chinese Hamster Cell Line

We describe here the derivation, characterization, and use of clonal cadmium-resistant (Cdr) strains of the Chinese hamster cell line CHO which differ in their metallothionein (MT) induction capacity. By nondenaturing polyacrylamide gel electrophoresis, we showed that the stable Cdr phenotype is correlated with the augmented expression of both isometallothioneins (MTI and MTII). In cells resistant to concentrations of CdCl2 exceeding 20 microM, coordinate amplification of genes encoding both isometallothioneins was demonstrated by using cDNA MT-coding sequence probes and probes specific for 3'-noncoding regions of Chinese hamster MTI and MTII genes. Molecular and in situ hybridization analyses supported close linkage of Chinese hamster MTI and MTII genes, which we have mapped previously to Chinese hamster chromosome 3. This suggests the existence of a functionally related MT gene cluster in this species. Amplified Cdr variants expressing abundant MT and their corresponding Cds parental CHO cells should be useful for future studies directed toward elucidating the mechanisms that regulate expression of the isometallothioneins.

scholarly journals PCR-RFLP of 16S ribosomal DNA to confirm the identification of Enterococcus gallinarum and Enterococcus casseliflavus isolated from clinical and food samples

2010 ◽  
Vol 43 (1) ◽  
pp. 100-101 ◽  
Author(s):  
Aline Weber Medeiros ◽  
Pedro d'Azevedo ◽  
Rebeca Inhoque Pereira ◽  
Ana Paula Cassenego ◽  
Sueli Van Der Sand ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Food Samples ◽  
Correct Identification ◽  
Accurate Identification ◽  
16S Ribosomal Dna ◽  
Pcr Rflp ◽  
Enterococcus Casseliflavus ◽  
Dna Pattern ◽  
Enterococcus Gallinarum ◽  
Rdna Analysis

INTRODUCTION: This study aimed to confirm the identification of Enterococcus gallinarum and Enterococcus casseliflavus isolated from clinical and food samples by PCR-RFLP. METHODS: Fifty-two strains identified by conventional biochemical exams were submitted to PCR amplification and digested with HinfI. Only 20 (38.5%) of the 52 strains showed a DNA pattern expected for E. gallinarum and E. casseliflavus. RESULTS: Analysis of the results of this study showed that E. gallinarum and E. casseliflavus are occasionally erroneously identified and confirmed the potential application of 16S rDNA analysis for accurate identification of these species. CONCLUSIONS: A correct identification is important to distinguish between intrinsic and acquired vancomycin resistance.

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