Polymorphism of the exon 3 of leptin gene in Malpura sheep

2016 ◽  
Author(s):  
A.S. Meena ◽  
A. Sahoo ◽  
R. S. Bhatt ◽  
A.S. Meena
Keyword(s):  
Genetic Variants ◽  
Genomic Dna ◽  
Extraction Method ◽  
Allelic Variation ◽  
Restriction Enzymes ◽  
Allelic Frequency ◽  
Adipose Tissues ◽  
Allelic Variants ◽  
Pcr Rflp ◽  
Pcr Products

Leptin (LEP) is primarily expressed in the adipose tissues. It regulates the feed intake, energy metabolism and body composition and plays a crucial role in regulating body weight and growth in mammals. The aim of the present study was to find out an allelic variation in leptin gene of Malpura sheep. A total of 112 Malpura sheep were selected and the genomic DNA was isolated by phenol-chloroform extraction method. PCR was carried out in order to amplify the 471 bp fragment of the exon 3 coding sequence of the leptin gene. The genotyping was done by PCR-RFLP technique. PCR products were digested with three (BcnI, SsiI and OliI) restriction enzymes. For detection of allelic variants, three non-synonymous SNPs were found in Malpura sheep. The A271G and A316C loci were found mono-morphic, while T387G locus was found polymorphic. Two genetic variants (G and T) and three genotypes (GG, GT and TT) were found in Malpura sheep. The allelic frequency of G and T allele at T387G locus was found 0.82 and 0.18, respectively.

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 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 Analysis of Slovak Spotted breed for bovine beta casein A1 variant as risk factor for human health.

1970 ◽  
Vol 60 (4) ◽  
Author(s):  
Martina Miluchová ◽  
Michal Gábor ◽  
Anna Trakovická
Keyword(s):  
Diabetes Mellitus ◽  
Genomic Dna ◽  
Total Population ◽  
Human Consumption ◽  
Effective Number ◽  
Immune Systems ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Beta Casein ◽  
Commercial Kit

The goal of work was identification A1 variant of bovine beta casein which involves ischemic heart disease and diabetes mellitus in human. The digestion of A1beta casein can result in the production of bioactive beta casomorphin-7 (BCM-7); this is not the case with A2. This bioactive peptide has been linked to physiological traits that may elicit effects on components of the vascular and immune systems. The material involved 111 Slovak Spotted breed. Bovine genomic DNA was extracted from whole blood by using commercial kit, and used in order to estimate beta-casein genotypes by means of PCR-RFLP method. The PCR products were digested with DdeI restriction enzyme. In the population included in the study were detected all three genotypes, homozygote genotype A1A1 (14 animals), heterozygote genotype A1A2 (37 animals) and homozygote genotype A2A2 (60 animals). In the total population of cattle homozygotes A2A2-0.5405 were the most frequent, while homozygotes A1A1-0.1261 were the least frequent ones. This suggests a superiority of allele A2 (0.7072) which does not produce BCM-7, and thus is safe for human consumption. The expected homozygosity for gene CSN2 is in the population stated a slight increase in homozygosity (0.5858). This caused a slight decrease in the level of possible variability realization (41.80%), which corresponds to the effective number of alleles (1.7071).

scholarly journals DISTINCTIONS AMONG ALLELIC VARIANTS ASSOCIATED WITH CHROMOSOME 3 INVERSIONS IN DROSOPHILA PSEUDOOBSCURA AND DROSOPHILA PERSIMILIS

Genetics ◽  
1980 ◽  
Vol 96 (3) ◽  
pp. 727-741
Author(s):  
R A Norman ◽  
Satya Prakash
Keyword(s):  
Acid Phosphatase ◽  
Genetic Variants ◽  
Allelic Variation ◽  
The Other ◽  
Drosophila Pseudoobscura ◽  
Chromosome 3 ◽  
Santa Cruz ◽  
Allelic Variants ◽  
The Common ◽  
Drosophila Persimilis

ABSTRACT Efforts were made to discriminate new genetic variants among electrophoretic alleles that are associated with chromosome 3 inversions of Drosophila pseudoobscura and D. persimilis. Apparent genetic similarities for electrophoretic alleles between these two species and among the common inversions they carry were reexamined by altering gel concentration and buffer pH. At the amylase locus, the 1.09 electrophoretic allele could be further separated into two allelic classes that differentiated the WT and KL arrangements. Similarly, the 0.84 electrophoretic allele was divided into two allelic classes, one characteristic of the Santa Cruz phylad arrangements, TL and SC, and the other found in strains of the Standard phylad arrangements and CH. Uncommon amylase alleles proved to be different alleles in the two species. No new allelic variants, however, could be found among strains with the amylase 1.00 allele, the commonest allele in the Standard phylad of both species. No major new allelic variation was detected for acid phosphatase-3 and larval protein-10 that revealed any further differentiation among species or inversions. Variation at all three loci in strains of the Bogota population remained genetically similar to variation in strains of mainland D. pseudoobscura.

scholarly journals Genetic Variants of k-Casein and β-Lactoglobulin Genes and Their Association with Protein and Milk Components of Holstein Friesian Cows at Small Farmers in Lembang, West Java

2017 ◽  
Vol 2 (6) ◽  
pp. 86 ◽  
Author(s):  
Anggraeni Anggraeni ◽  
A. Anneke ◽  
H.S. Nury ◽  
E. Andreas ◽  
C. Sumantri
Keyword(s):  
Genetic Variants ◽  
Milk Protein ◽  
Restriction Enzymes ◽  
Dairy Farmers ◽  
West Java ◽  
Holstein Friesian ◽  
Pcr Rflp ◽  
Milk Components ◽  
Milk Component ◽  
Β Lactoglobulin

Genetic variants of CSN3 and LGB genes and their effects on protein and milk components were studied in Holstein Friesian at small dairy farmers in Lembang District, West Java, Indonesia.  Allelic variants were identified by PCR-RFLP technique using restriction enzymes of Pst I for the CSN3 gene and Hae III for the LGB gene.  The CSN3 gene was dominated by AB genotype.   Milk protein was not affected by genotypes of the two genes.  Only fat content was significantly affected (P <0.05) by the CSN3 gene with AB cows having the highest fat to AA and BB cows (3.76% vs. 3.26% and 3.34%). Keywords: CSN3 gene; LGB gene; milk protein; and milk component.

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 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. .

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.

Rapid and reliable identification of rice genomes by RFLP analysis of PCR-amplified Adh genes

Genome ◽  
2001 ◽  
Vol 44 (6) ◽  
pp. 1136-1142 ◽  
Author(s):  
Song Ge ◽  
Tao Sang ◽  
Bao-rong Lu ◽  
De-yuan Hong
Keyword(s):  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Pcr Primers ◽  
Specific Pcr ◽  
Adh Genes ◽  
Pcr Rflp ◽  
Restriction Sites ◽  
Pcr Products ◽  
Adh Gene ◽  
Genus Oryza

The rice genus (Oryza L.) consists of 24 species with 10 recognized genome types. With the realization of many useful genes in species of wild rice, continuous efforts have been made to understand their genomic composition and relationships. However, the identification of rice genomes has often been difficult owing to complex morphological variation and formation of allotetraploids. Here we propose a rapid and reliable method for identifying rice genomes based on the restriction sites of PCR-amplified Adh genes. The experimental procedure was as follows: (i) amplify a portion of Adh1 and Adh2 genes with the locus-specific PCR primers; (ii) digest PCR products with restriction enzymes that distinguish different genomes; and (iii) run the digested products on 1.4% agarose gel, and photograph. Using various combinations of restriction digestion of the two Adh genes, all of the rice genomes can be identified.Key words: Adh gene, genome, identification, Oryza L., PCR–RFLP.

scholarly journals Identification and differentiation among chicken’s, duck’s, quail’s, rabbit’s and turkey's meat using PCR-RFLP technique

2015 ◽  
Vol 31 (1) ◽  
pp. 101-108 ◽  
Author(s):  
S.M. Abdel-Rahman ◽  
A.M. Elmaghraby ◽  
A.S. Haggag
Keyword(s):  
Mitochondrial Dna ◽  
Cytochrome B ◽  
Fragment Length ◽  
Restriction Analysis ◽  
Restriction Enzymes ◽  
Pcr Product ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Species Specific ◽  
Rflp Assay

PCR-RFLP technique was developed for identification and differentiation among chicken?s, duck?s, quail?s, rabbit?s and turkey's meat. DNA from small amount of muscles (0.05 g) was extracted and a region of mitochondrial DNA (cytochrome-b gene) in chicken, duck, quail, rabbit and turkey was amplified by PCR. Fragment length of the PCR product was 371 bp in chicken, 374 bp in duck and rabbit and 377 bp in both quail and turkey. Six nucleotides different makes it difficult to differentiate among these five species-specific meat. For differentiation, three different restriction enzymes (DdeI, MspI and TaqI) were used to digest the PCR products. Restriction analysis showed difference among chicken?s, duck?s, quail?s, rabbit?s and turkey's meat. Where, DdeI yielded two fragments (291 and 83 bp) only in rabbit?s meat. MspI yielded three fragments (221, 85 and 65 bp) in chicken?s meat and two fragments (290 and 87 bp) in both quail?s and turkey's meat. TaqI yielded three fragments (146, 134 and 94 bp) in duck?s meat and two fragments (226 and 151 bp) in quail?s meat. The use of Cytb- PCR-RFLP assay allowed a direct and fast authentication and differentiation among chicken?s, duck?s, quail?s, rabbit?s and turkey's meat.

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