Detection of A1 and A2 Alleles at Beta-casein Locus in Bargur and Umblachery (Indian Zebu) Cattle Breeds by Allele-specific PCR

Indian Journal of Animal Research ◽

10.18805/ijar.b-4273 ◽

2021 ◽

Author(s):

A. Raja ◽

R. Rajendran ◽

P. Ganapathi

Keyword(s):

Information Content ◽

Polymorphism Information Content ◽

Zebu Cattle ◽

Effective Number ◽

Cattle Breeds ◽

Specific Pcr ◽

Genotype Frequencies ◽

Beta Casein ◽

Allele Specific ◽

Allele Specific Pcr

Background: Many genetic variants of beta-casein in different breeds of cattle have been reported. The A1 and A2 are the most common variants. The breeds of Zebu cattle have high frequency of A2 allele or monomorphic for A2 allele. The current study aimed to screen Indian Zebu cattle breeds, Bargur and Umblachery, for A1 and A2 alleles at beta-casein locus.Methods: A total of 48 Bargur and 42 Umblachery cattle were genotyped for β-casein (CSN2) gene using allele-specific PCR. The gene and genotype frequencies were estimated. The theoretical heterozygosity (Heexp), experimental heterozygosity (Heobs), polymorphism information content (PIC), expected homozygosity (E), effective number of alleles (ENA) and level of possible variability realization (V%) were calculated.Result: The investigation revealed the presence of both A1 and A2 alleles at beta-casein locus in both Bargur and Umblachery cattle breeds. The A1A1 genotype was not observed in both the breeds. The frequencies of A1A2 and A2A2 genotypes were 0.125 and 0.875 respectively in Bargur and 0.050 and 0.950 respectively in Umblachery breed. The study indicated the predominance of A2 variant in both the breeds. The frequencies of A1 and A2 alleles were 0.063 and 0.937 respectively in Bargur and 0.02 and 0.98 respectively in Umblachery breed. The values of experimental heterozygosity (Heobs), theoretical heterozygosity (Heexp), polymorphism information content (PIC), expected homozygosity (E), effective number of alleles (ENA), level of possible variability realization (V%) were 0.125, 0.1163, 0.1095,0.8837, 1.131 and 11.88 respectively in Bargur breed. These values were 0.048, 0.0468, 0.0458, 0.9532, 1.049 and 4.79 respectively in Umblachery population. The observed heterozygosity and PIC values revealed the existence of very low genetic variability in the tested populations. The present work will be a contribution to the study on beta-casein locus in Indian zebu cattle.

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Evaluation of b-casein variants in Egyptian goat, sheep and cattle by allele specific PCR and relevance to b- casomorphin

Indian Journal of Animal Research ◽

10.18805/ijar.b-805 ◽

2018 ◽

Author(s):

Ahmed M. Darwish ◽

Ghada H. H. El Nady ◽

Neama I. Ali ◽

Aly Z. E. Abdelsalam

Keyword(s):

Polymerase Chain Reaction ◽

Genetic Variants ◽

Chain Reaction ◽

Specific Polymerase Chain Reaction ◽

Specific Pcr ◽

Sheep Milk ◽

Beta Casein ◽

Allele Specific ◽

Polymerase Chain ◽

Allele Specific Pcr

The beta casein gene (CSNS2) has 12 genetic variants divided into two groups: the first group (A1, B, and G) which differ from the second group (A2, A3 C, D, E, F, H, H2 and I) where A base replaces C base, this leads to potential liberation of a bioactive peptide, b-casomorphin, upon digestion where a histidine replaces a proline at position 67. The allele specific polymerase chain reaction (AS-PCR) was evaluated to distinguish between the beta casomorphin releasing variants (A1 and B) and the non-releasing variants. The sequence analysis was used to determine these variants and confirm it in goat, sheep and cattle. The results showed that cattle carrying allele A1 either homozygous or heterozygous more than sheep and goats. The allele frequency of A1 and A2 is 0.44, 0.56 in goats, 0.43, 0.57 in sheep and 0.54, 0.46 in cattle, respectively. The sequence results reported changing of C base to A base in goat, sheep and cattle. Therefore, this study reported that goat and sheep milk was more safe than cattle milk.

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

Thrombosis and Haemostasis ◽

10.1055/s-0038-1650362 ◽

1996 ◽

Vol 75 (05) ◽

pp. 757-759 ◽

Cited By ~ 11

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.

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MUPrimer : a tool for finding allele specific PCR-primers for homologous gene sequences

10.32469/10355/6660 ◽

2009 ◽

Author(s):

M. Mursaleen Ahmad

Keyword(s):

Pcr Primers ◽

Homologous Gene ◽

Gene Sequences ◽

Specific Pcr ◽

Allele Specific ◽

Allele Specific Pcr

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Development of genic KASP SNP markers from RNA-Seq data for map-based cloning and marker-assisted selection in maize

BMC Plant Biology ◽

10.1186/s12870-021-02932-8 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Zhengjie Chen ◽

Dengguo Tang ◽

Jixing Ni ◽

Peng Li ◽

Le Wang ◽

...

Keyword(s):

Marker Assisted Selection ◽

Inbred Lines ◽

Average Density ◽

Snp Markers ◽

Data Sets ◽

Rna Seq ◽

Specific Pcr ◽

Maize Inbred Lines ◽

Allele Specific ◽

Allele Specific Pcr

Abstract Background Maize is one of the most important field crops in the world. Most of the key agronomic traits, including yield traits and plant architecture traits, are quantitative. Fine mapping of genes/ quantitative trait loci (QTL) influencing a key trait is essential for marker-assisted selection (MAS) in maize breeding. However, the SNP markers with high density and high polymorphism are lacking, especially kompetitive allele specific PCR (KASP) SNP markers that can be used for automatic genotyping. To date, a large volume of sequencing data has been produced by the next generation sequencing technology, which provides a good pool of SNP loci for development of SNP markers. In this study, we carried out a multi-step screening method to identify kompetitive allele specific PCR (KASP) SNP markers based on the RNA-Seq data sets of 368 maize inbred lines. Results A total of 2,948,985 SNPs were identified in the high-throughput RNA-Seq data sets with the average density of 1.4 SNP/kb. Of these, 71,311 KASP SNP markers (the average density of 34 KASP SNP/Mb) were developed based on the strict criteria: unique genomic region, bi-allelic, polymorphism information content (PIC) value ≥0.4, and conserved primer sequences, and were mapped on 16,161 genes. These 16,161 genes were annotated to 52 gene ontology (GO) terms, including most of primary and secondary metabolic pathways. Subsequently, the 50 KASP SNP markers with the PIC values ranging from 0.14 to 0.5 in 368 RNA-Seq data sets and with polymorphism between the maize inbred lines 1212 and B73 in in silico analysis were selected to experimentally validate the accuracy and polymorphism of SNPs, resulted in 46 SNPs (92.00%) showed polymorphism between the maize inbred lines 1212 and B73. Moreover, these 46 polymorphic SNPs were utilized to genotype the other 20 maize inbred lines, with all 46 SNPs showing polymorphism in the 20 maize inbred lines, and the PIC value of each SNP was 0.11 to 0.50 with an average of 0.35. The results suggested that the KASP SNP markers developed in this study were accurate and polymorphic. Conclusions These high-density polymorphic KASP SNP markers will be a valuable resource for map-based cloning of QTL/genes and marker-assisted selection in maize. Furthermore, the method used to develop SNP markers in maize can also be applied in other species.

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Two User-Friendly Molecular Markers Developed for the Identification of Hybrid Lethality Genes in Brassica oleracea

Agronomy ◽

10.3390/agronomy11050982 ◽

2021 ◽

Vol 11 (5) ◽

pp. 982

Author(s):

Zhiliang Xiao ◽

Congcong Kong ◽

Fengqing Han ◽

Limei Yang ◽

Mu Zhuang ◽

...

Keyword(s):

Molecular Markers ◽

Brassica Oleracea ◽

Rapid Identification ◽

Inbred Lines ◽

Hybrid Lethality ◽

Specific Pcr ◽

Allele Specific ◽

User Friendly ◽

Allele Specific Pcr ◽

Kasp Marker

Cabbage (Brassica oleracea) is an important vegetable crop that is cultivated worldwide. Previously, we reported the identification of two dominant complementary hybrid lethality (HL) genes in cabbage that could result in the death of hybrids. To avoid such losses in the breeding process, we attempted to develop molecular markers to identify HL lines. Among 54 previous mapping markers closely linked to BoHL1 or BoHL2, only six markers for BoHL2 were available in eight cabbage lines (two BoHL1 lines; three BoHL2 lines; three lines without BoHL); however, they were neither universal nor user-friendly in more inbred lines. To develop more accurate markers, these cabbage lines were resequenced at an ~20× depth to obtain more nucleotide variations in the mapping regions. Then, an InDel in BoHL1 and a single-nucleotide polymorphism (SNP) in BoHL2 were identified, and the corresponding InDel marker MBoHL1 and the competitive allele-specific PCR (KASP) marker KBoHL2 were developed and showed 100% accuracy in eight inbred lines. Moreover, we identified 138 cabbage lines using the two markers, among which one inbred line carried BoHL1 and 11 inbred lines carried BoHL2. All of the lethal line genotypes obtained with the two markers matched the phenotype. Two markers were highly reliable for the rapid identification of HL genes in cabbage.

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