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

2019 ◽  
Author(s):  
Ahmed Abdel-kader 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, 651bp 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 Comparative analysis of IGFBP-3 gene sequence in Egyptian sheep, cattle, and buffalo

2019 ◽  
Author(s):  
Ahmed Abdel-kader 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, 651bp 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 Comparative analysis of IGFBP-3 gene sequence in Egyptian sheep, cattle, and buffalo

2019 ◽  
Author(s):  
Ahmed Abdel-kader 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, 651bp 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 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.

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.

IDENTIFYING THE C677T POLYMORPHISM OF MTHFR GENE BY PCR-RFLP TECHNIQUE

2011 ◽  
pp. 28-35
Author(s):  
Keyword(s):  
Internal Control ◽  
Restriction Site ◽  
Mthfr Gene ◽  
C677t Polymorphism ◽  
Mthfr Genotype ◽  
Restriction Digest ◽  
Genotype Frequencies ◽  
Pcr Rflp ◽  
Sequencing Technique ◽  
Volunteer Group

Background: The C677T polymorphism of MTHFR gene is a risk factor of many diseases. This study is aimed at: (1) Improving a PCR-RFLP process with the own designed primers to identify the C677T polymorphism of MTHFR gene. (2) Evaluating the prevalence of the C677T polymorphism of MTHFR gene in volunteer group. Materials and method: DNA samples was extracted from peripheral blood of 60 volunteers. Designing primers by using FastPCR software, then improving PCR technique. Standardizing the optimal conditions of restriction digest by HinfI. Confirming the results of polymorphism by DNA sequencing technique. Results: We designed successfully primers to amplify fragment of MTHFR gene including C677T polymorphism and an obligatory restriction site of HinfI (as internal control). 0.5 µl of HinfI enzyme (10 U/µl) is enough for restriction digest. The MTHFR genotype frequencies were: 71.67 % (677CC); 25% (677CT); and 3.33 % (677TT). Conclusion: We standardized successfully PCR-RFLP technique to identifying C677T polymorphism of MTHFR gene. Keywords: C677T polymorphism, MTHFR gene, PCR-RFLP

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.

Association Study of Fecundity Gene BMPR1B with Prolificacy in Surti Goats under Farm and Field Condition

2019 ◽  
Vol 14 (04) ◽  
Author(s):  
N. S. Dangar ◽  
G. M. Pandya ◽  
U. V. Ramani ◽  
Y. D. Padheriya ◽  
T. Sangma ◽  
...  
Keyword(s):  
Litter Size ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Pcr Primers ◽  
Dual Purpose ◽  
Protein Receptor ◽  
Morphogenetic Protein ◽  
Pcr Rflp ◽  
Base Size ◽  
Mutation Information

The Surti is a dual purpose goat breed of Gujarat. The bone morphogenetic protein receptor type 1B (BMPR1B) gene of transforming growth factor beta (TGF-β) superfamily ligands is playing a role in ovulation as well as litter size. Mutation in Exon-6 region of BMPR1B gene with base size 190 bp reported increasing litter size. Based on the known mutation information in goat and sheep, PCR primers were designed to screen polymorphism in total 100 Surti goats, 50 Surti goats from University Farm, Navsari and 50 Surti goats from field units of Southern part of Gujarat. During PCR-RFLP study no polymorphic sites were found for Exon-6 region of BMPR1B on Surti goats. Moreover, the twinning rate was 10% in first parity and higher in subsequent second (62.5%) and third (76.8%) parties.

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