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 Fingerprinting of fecB gene in five Egyptian sheep breeds

2009 ◽  
Vol 25 (3-4) ◽  
pp. 205-212 ◽  
Author(s):  
Hanafy el ◽  
Saadani el
Keyword(s):  
Point Mutation ◽  
Restriction Enzyme ◽  
Base Pair ◽  
Genomic Dna ◽  
Restriction Site ◽  
Wild Type ◽  
Specific Primer ◽  
Sheep Breeds ◽  
Pcr Rflp ◽  
Pcr Products

Recently, many aspects of FecB gene, including reproductive endocrinology, organs development and body mass have been studied. FecB has an additive effect on litter size and ovulation. The present investigation was carried out to study polymorphism by forced PCR-RFLP of FecB gene in five Egyptian local sheep breeds and its comparison with other foreign sheep breeds. Genomic DNA was isolated from a total of 100 animals of Egyptian sheep breeds namely Rahmani, Ossimi, Awassi, Barki and Awassi x Barki crossbred. Forced PCR of the FecB gene 190 base pair (bp) was amplified using specific primer designed to introduc a point mutation in the resulting PCR products with FecB carrier sheep containing an AvaII restriction site (G|GACC), whereas products from noncarriers lacked (of) this site. Digestion of FecB gene 190 base pair with AvaII restriction enzyme resulted in non carrier 190 bp band (wild type) in all the animals belonging to the five Egyptian breeds studied revealing absence of this restriction site in those five breeds. .

scholarly journals Genetic diversity of <i>κ</i>-casein (<i>CSN3</i>) and lactoferrin (<i>LTF</i>) genes in the endangered Turkish donkey (<i>Equus asinus</i>) populations

2019 ◽  
Vol 62 (1) ◽  
pp. 77-82 ◽  
Author(s):  
Fulya Özdil ◽  
Hasan Bulut ◽  
Raziye Işık
Keyword(s):  
Restriction Enzyme ◽  
Animal Species ◽  
Restriction Enzymes ◽  
Production Traits ◽  
Single Nucleotide ◽  
Equus Asinus ◽  
Pcr Rflp ◽  
Hardy Weinberg Equilibrium ◽  
Polymerase Chain ◽  
Enzyme Recognition

Abstract. In this study, the κ-casein (CSN3) and lactoferrin (LTF) genes which were found in association with milk production traits in different animal species were studied firstly in Turkish donkey populations. A total of 108 donkeys from different regions of Turkey were used in order to reveal the different genotypes of CSN3 and LTF genes by using polymerase chain reaction – restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing methods. To determine the genetic polymorphism, we attempted to digest a fragment of 235 bp of the CSN3 gene and a fragment of 751 bp of the LTF gene using PstI, and DraII, EagI and MboI restriction enzymes, respectively. Neither the CSN3 gene nor the LTF gene had enzyme recognition sites with the PstI, DraII and MboI restriction enzymes in all of the studied samples. However, the LTF gene was only distinguished with the EagI restriction enzyme. Three genotypes were identified in the LTF gene with the EagI restriction enzyme: GG homozygotes (667, 84 bp), AG heterozygotes (751; 667, 84 bp) and AA homozygotes (751 bp). The transition from guanine to adenine in 89 bp of the LTF gene lacks the restriction site and different genotypes are obtained. This novel single nucleotide polymorphism (SNP) has been firstly detected in donkeys. According to the results, the G allele was predominant in the LTF-EagI gene in the studied Turkish donkey populations. In this study, all the genotype distributions of LTF-EagI were not found in Hardy–Weinberg equilibrium (P<0.05). The CSN3 and LTF genes have not been studied before in donkeys, so the results are the preliminary results of these gene regions in donkeys.

scholarly journals Rapid differentiation of Staphylococcus aureus isolates harbouring egc loci with pseudogenes ψent1 and ψent2 and the selu or selu v gene using PCR-RFLP

2007 ◽  
Vol 56 (2) ◽  
pp. 208-216 ◽  
Author(s):  
Mark M. Collery ◽  
Cyril J. Smyth
Keyword(s):  
Dna Sequencing ◽  
Restriction Enzyme ◽  
Sequence Similarity ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Restriction Enzyme Digestion ◽  
Pcr Rflp ◽  
Intergenic Regions ◽  
Unique Restriction ◽  
Genomic Regions

The egc locus of Staphylococus aureus harbours two enterotoxin genes (seg and sei) and three enterotoxin-like genes (selm, seln and selo). Between the sei and seln genes are located two pseudogenes, ψent1 and ψent2, or the selu or selu v gene. While these two alternative sei–seln intergenic regions can be distinguished by PCR, to date, DNA sequencing has been the only confirmatory option because of the very high degree of sequence similarity between egc loci bearing the pseudogenes and the selu or selu v gene. In silico restriction enzyme digestion of genomic regions encompassing the egc locus from the 3′ end of the sei gene through the 5′ first quarter of the seln gene allowed pseudogene- and selu- or selu v-bearing egc loci to be distinguished by PCR-RFLP. Experimental application of these findings demonstrated that endonuclease HindIII cleaved PCR amplimers bearing pseudogenes but not those with a selu or selu v gene, while selu- or selu v-bearing amplimers were susceptible to cleavage by endonuclease HphI, but not by endonuclease HindIII. The restriction enzyme BccI cleaved selu- or selu v-harbouring amplimers at a unique restriction site created by their signature 15 bp insertion compared with pseudogene-bearing amplimers, thereby allowing distinction of these egc loci. PCR-RFLP analysis using these restriction enzymes provides a rapid, easy to interpret alternative to DNA sequencing for verification of PCR findings on the nature of an egc locus type, and can also be used for the primary identification of the intergenic sei–seln egc locus type.

Restriction enzyme mapping of the nuclear ribosomal cistron in selected Laminariales (Phaeophyta): a phylogenetic assessment

1991 ◽  
Vol 69 (12) ◽  
pp. 2647-2654 ◽  
Author(s):  
G. W. Saunders ◽  
L. D. Druehl
Keyword(s):  
Key Words ◽  
Restriction Enzyme ◽  
Restriction Site ◽  
Intergenic Spacer ◽  
Restriction Maps ◽  
Restriction Enzyme Mapping ◽  
Eisenia Arborea ◽  
Macrocystis Integrifolia ◽  
Enzyme Mapping ◽  
Ribosomal Cistron

Restriction enzyme mapping of the nuclear ribosomal cistron was completed for a variety of Laminariales. Taxa investigated included Alaria marginata Postels and Ruprecht, Egregia menziesii (Turner) Areschoug, Eisenia arborea Areschoug, Lessoniopsis littoralis (Tilden) Reinke, Macrocystis integrifolia Bory, Nereocystis leutkeana (Mertens) Postels and Ruprecht, Postelsia palmaeformis Ruprecht, and Pterygophora californica Ruprecht, with Sargassum muticum (Yendo) Fensholt (Fucales) as an outgroup. The restriction maps establish a foundation for future phylogenetic, as well as other molecular, investigations in the kelp. We also assess restriction enzyme mapping of the nuclear ribosomal cistron for suitability in resolving intrafamilial and interfamilial taxonomic relationships in the Laminariales. The intergenic spacer proved too variable to be of use for phylogenetic comparisons at this level. Conversely, the gene regions were highly conserved, with only three restriction-site differences observed among all the laminarialean taxa investigated. Key words: Laminariales, rRNA, restriction enzyme mapping.

Detection of Theileria annulata by the PCR-RFLP in ticks (Acari, Ixodidae) collected from cattle in West and North-West Iran

2011 ◽  
Vol 56 (1) ◽  
Author(s):  
Mosa Tavassoli ◽  
Mohammad Tabatabaei ◽  
Bijan Nejad ◽  
Mehran Tabatabaei ◽  
Amin Najafabadi ◽  
...  
Keyword(s):  
Restriction Enzymes ◽  
Theileria Annulata ◽  
Gene Encoding ◽  
Hyalomma Anatolicum Anatolicum ◽  
North West ◽  
Blood Smears ◽  
Pcr Rflp ◽  
Single Pattern ◽  
Pcr Products ◽  
Bovine Theileriosis

AbstractThe presence of potential vectors, ticks, and susceptible hosts of bovine malignant theileriosis in all parts of Iran pose a real threat to food animal industry. The present study was conducted to determine the infection rate of ticks collected from naturally occurring bovine theileriosis in West and North-West Iran. Two hundred and thirty seven cattle suspected of suffering from theileriosis were investigated for the presence of Theileria annulata in the blood smears and any tick species on their body. In this study, 402 ticks were obtained from 99 cattle. The examination of 402 ticks by polymerase chain reaction (PCR) using primers derived from the gene encoding heat shock protein70 (Hsp70) revealed that 39.9% of Hyalomma anatolicum anatolicum, 3.5% of H. asiaticum asiaticum, and 18.2% H. anatolicum excavatum, were infected with T. annulata. The results suggest that H. a. anatolicum may play a major role in transmission of T. annulata infection in Iran. Finally, digestion of the PCR products of T. annulata with two different restriction enzymes produced only a single pattern.

scholarly journals Profile of follicle-stimulating hormone and polymorphism of follicle-stimulating hormone receptor in Madrasin cattle with ovarian hypofunction

2020 ◽  
Vol 13 (5) ◽  
pp. 879-883
Author(s):  
Budi Utomo ◽  
Emmanuel Djoko Putranto ◽  
Amaq Fadholly
Keyword(s):  
Fragment Length Polymorphism ◽  
Follicle Stimulating Hormone ◽  
Restriction Enzymes ◽  
Chain Reaction ◽  
Blood Samples ◽  
Fshr Gene ◽  
Polymerase Chain ◽  
Genetic Makeup ◽  
Restriction Fragment Length ◽  
Stimulating Hormone

Background and Aim: The follicle-stimulating hormone (FSH) gene is an essential regulator of fertility in livestock. This study aims to provide information on the genetic makeup of Madrasin cattle experiencing hypofunction by the FSH profile and FSH receptors (FSHR) polymorphism. Materials and Methods: Blood samples were collected from the Bangkalan regency in Indonesia. DNA was isolated and purified following the extraction protocol of polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism. Results: Our results showed that the FSH gene had a band length of 310 bp and produce two alleles (A and B) with restriction enzymes at 250 bp, 230 bp, and 145 bp. Furthermore, the FSHR gene had a band length of 303 bp and produced two homozygous genotypes: GG at bp 239 and CC at bp 188. Conclusion: Based on these differences, there was no change in allele frequency and genotype between Madura and Madrasin cattle due to crossbreeding with Limousin cattle. Thus, further detailed investigations of Madrasin cattle are required to elucidate the profile of the LH and LHR genes.

scholarly journals Survey of mutations in prolificacy genes in Santa Ines and Morada Nova sheep

2017 ◽  
Vol 69 (4) ◽  
pp. 1047-1053
Author(s):  
G.M.L. Holanda ◽  
J.C. Oliveira ◽  
D.M.F. Silva ◽  
S.S.N. Rocha ◽  
V. Pandolfi ◽  
...  
Keyword(s):  
Restriction Site ◽  
Fragment Length Polymorphism ◽  
Nucleotide Sequencing ◽  
Chain Reaction ◽  
Specific Primers ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Santa Inês ◽  
Restriction Fragment Length

ABSTRACT Polymorphisms in the BMP-15 gene related to Galway (FecXG) and Inverdale (FecXI) and in the BMPR-1B gene known as Booroola (FecB) mutations were investigated using the Polymerase Chain Reaction - Restriction Fragment Length Polymorphism (PCR-RFLP) method, on sheep from the breeds Santa Inês (n= 574) and Morada Nova (n=282). DNA was extracted and amplified through PCR with specific primers that introduced a restriction site in association with the mutation. The PCR products were submitted to endonucleases. The experiment found no FecXG and FecXI mutations. Six samples of animals with multiple offspring/birth history presented polymorphism for FecB similar to control samples, but this pattern was not confirmed by nucleotide sequencing. Although the absence of these mutations in the studied breeds, other factors related to prolificacy should be investigated to explain the inherent prolificity mechanisms.

PCR-RFLP ANALYSIS OF THE CHLOROPLAST GENE trn K IN THE PINACEAE, WITH SPECIAL REFERENCE TO THE SYSTEMATIC POSITION OF CATHAYA

1998 ◽  
Vol 46 (4) ◽  
pp. 265-271 ◽  
Author(s):  
Xiao-Quan Wang ◽  
Ying Han ◽  
De-yuan Hong
Keyword(s):  
Molecular Phylogeny ◽  
Restriction Site ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Systematic Position ◽  
Chloroplast Gene ◽  
Parsimony Method ◽  
Pcr Rflp ◽  
Restriction Sites ◽  
Dollo Parsimony

The molecular phylogeny of the Pinaceae represented by 13 species of 10 genera was constructed from PCR-RFLP analysis of the chloroplast gene trn K, which was approximately 2557 bp long. Ninety-two restriction sites, of which 68 were variable, were identified by 16 restriction enzymes. Thirty-five of the 68 polymorphic sites were phylogenetically informative. The restriction site data were analyzed by PAUP (version 3.1.1) with both the Wagner parsimony method and the Dollo parsimony method. As a result, Dollo and Wagner parsimonious trees have similar topologies except for the position of Cedrus. The Abies-Keteleeria-Tsuga-Pseudolarix clade was well resolved in all trees. Pseudotsuga is closely related to Larix, while Abies is relatively closely related to Keteleeria. As an isolated genus, Cathaya is distantly related to the Abies-Keteleeria-Tsuga-Pseudolarix clade, and is not very closely related to any other genus of the Pinaceae.

Diversity of the desert truffle Terfezia boudieri Chatin. in southern Tunisia

2011 ◽  
Vol 57 (7) ◽  
pp. 599-605 ◽  
Author(s):  
Imed Sbissi ◽  
Faten Ghodhbane-Gtari ◽  
Mohamed Neffati ◽  
Hadda Ouzari ◽  
Abdellatif Boudabous ◽  
...  
Keyword(s):  
Its Region ◽  
Restriction Enzymes ◽  
Low Ph ◽  
Fruiting Bodies ◽  
Desert Truffle ◽  
Southern Tunisia ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Novel Taxa ◽  
Terfezia Boudieri

This study reports the genetic diversity of Terfezia boudieri collected from southern Tunisia. The study was carried out using 135 truffle fruiting bodies harvested from seven different locations. Twenty-eight Terfezia claveryi fruiting bodies were also sampled from one of the seven locations. A PCR-based technique was used to amplify the internal transcribed spacer (ITS) region of the rDNA, including the ITS1–5.8S–ITS2. The PCR products were digested with the four restriction enzymes RsaI, HhaI, AluI, and HinfI. Based on the HinfI patterns, T. boudieri specimens were separated into two different haplotypes (I and II). Nucleotide sequences of some representative amplicons were also obtained. Based on the phylogenetic results, three T. boudieri genotypes could be differentiated. One sequence, SKtb1, retrieved from PCR–RFLP of haplotype I, was obtained from a low pH soil in association with Helianthemum kahiricum . Based on the results presented in the current study, this isolate may represent a novel taxa within the Terfezia genus.

scholarly journals Derived Polymorphic Amplified Cleaved Sequence (dPACS): A Novel PCR-RFLP Procedure for Detecting Known Single Nucleotide and Deletion–Insertion Polymorphisms

2019 ◽  
Vol 20 (13) ◽  
pp. 3193 ◽  
Author(s):  
Shiv Shankhar Kaundun ◽  
Elisabetta Marchegiani ◽  
Sarah-Jane Hutchings ◽  
Ken Baker
Keyword(s):  
Gel Electrophoresis ◽  
Restriction Site ◽  
Restriction Enzymes ◽  
Nucleotide Polymorphisms ◽  
Acetyl Coa Carboxylase ◽  
Wild Type ◽  
Single Nucleotide ◽  
Pcr Rflp ◽  
Broadleaf Species ◽  
Selection Of

Most methods developed for detecting known single nucleotide polymorphisms (SNP) and deletion–insertion polymorphisms (DIP) are dependent on sequence conservation around the SNP/DIP and are therefore not suitable for application to heterogeneous organisms. Here we describe a novel, versatile and simple PCR-RFLP procedure baptised ‘derived Polymorphic Amplified Cleaved Sequence’ (dPACS) for genotyping individual samples. The notable advantage of the method is that it employs a pair of primers that cover the entire fragment to be amplified except for one or few diagnostic bases around the SNP/DIP being investigated. As such, it provides greater opportunities to introduce mismatches in one or both of the 35–55 bp primers for creating a restriction site that unambiguously differentiates wild from mutant sequences following PCR-RFLP and horizontal MetaPhorTM gel electrophoresis. Selection of effective restriction enzymes and primers is aided by the newly developed dPACS 1.0 software. The highly transferable dPACS procedure is exemplified here with the positive detection (in up to 24 grass and broadleaf species tested) of wild type proline106 of 5-enolpyruvylshikimate-3-phosphate synthase and its serine, threonine and alanine variants that confer resistance to glyphosate, and serine264 and isoleucine2041 which are key target-site determinants for weed sensitivities to some photosystem II and acetyl-CoA carboxylase inhibiting herbicides, respectively.

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