PCR Based Genotyping of Lulu Cattle of Nepal for A1, A2 Type Beta-caseins

Lulu is an indigenous breed of cattle (Bos taurus) found in high altitude regions of western Nepal. Population of Lulu cattle has been declining due to introgression with other exotic breeds to increase milk productivity. Here we aimed at finding potential approach for conserving Lulu cattle and its...

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Main Authors: Neena Amatya Gorkhali, Chhiring Sherpa, Nabina Budhathoki, Sudarshan GC, Srijana Lama, Prashanna Pokharel, Bhoj Raj Pokhrel, Saroj Sapkota
Format: Article
Language:English
Published: Nepal Agricultural Research Council 2020-03-01
Series:Journal of Nepal Agricultural Research Council
Subjects:
Online Access:https://www.nepjol.info/index.php/JNARC/article/view/28115
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author Neena Amatya Gorkhali
Chhiring Sherpa
Nabina Budhathoki
Sudarshan GC
Srijana Lama
Prashanna Pokharel
Bhoj Raj Pokhrel
Saroj Sapkota
author_facet Neena Amatya Gorkhali
Chhiring Sherpa
Nabina Budhathoki
Sudarshan GC
Srijana Lama
Prashanna Pokharel
Bhoj Raj Pokhrel
Saroj Sapkota
author_sort Neena Amatya Gorkhali
collection DOAJ
description Lulu is an indigenous breed of cattle (Bos taurus) found in high altitude regions of western Nepal. Population of Lulu cattle has been declining due to introgression with other exotic breeds to increase milk productivity. Here we aimed at finding potential approach for conserving Lulu cattle and its assets by studying the milk contents and investigating which variant of beta-casein protein is present in this breed. Beta caseins are an abundant protein in cow milk with A1 and A2 being the most common genetic variants of this protein. Consumption of A1 type of milk has numerous health-related complications whereas A2 type of milk has numerous human health promoting factors. We used restriction fragment length polymorphism (RFLP) for determining the A1 and A2 variant of beta casein in Lulu cattle. For performing DNA extraction, we collected (n = 18) blood samples of Lulu from Mustang and (n=17) Nepal Agriculture research council farm. The amplified fragments in 3% agarose at 251bp and 213bp respectively confirmed the presence of both A1 and A2 gene in Lulu; however, A2 was of greater abundance. Our study indicated that Lulu has A2 variant of beta-casein predominantly. The gene frequency of A1A1 is 0, A1A2 is 0.06 and A2A2 is 0.94. We further found that the allele frequency of A1 and A2 is 0.03 and 0.97 respectively. We designed special primer for sequencing CSN2 genes since A2 type beta casein gene was predominantly seen on Lulu. The sequencing result further supports our RFLP result as most of our samples have “C” nucleotide SNP in amplified CSN2 gene sequence. The Chi-square value of the current study is 0.04 which supports Hardy-Weinberg equilibrium inferring that Lulu cattle are still in the pure state, where there is no genetic introgression with the exotic breed for the sake of improvement of productivity.
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spelling doaj.art-c5a67361a47a46e186c76c75b0fcc5fa2022-12-21T18:02:06ZengNepal Agricultural Research CouncilJournal of Nepal Agricultural Research Council2392-45352392-45432020-03-0161566110.3126/jnarc.v6i0.28115PCR Based Genotyping of Lulu Cattle of Nepal for A1, A2 Type Beta-caseinsNeena Amatya Gorkhali0Chhiring Sherpa1Nabina Budhathoki2Sudarshan GC3Srijana Lama4Prashanna Pokharel5Bhoj Raj Pokhrel6Saroj Sapkota7Animal Breeding Division, Nepal Agricultural Research Council, Lalitpur, NepalSANN International College, Purbanchal University, Gairidhara, KathmanduSANN International College, Purbanchal University, Gairidhara, KathmanduSANN International College, Purbanchal University, Gairidhara, KathmanduSANN International College, Purbanchal University, Gairidhara, KathmanduUniversity of Alberta, Edmonton, Alberta, CanadaAnimal Breeding Division, Nepal Agricultural Research Council, Lalitpur, NepalAnimal Breeding Division, Nepal Agricultural Research Council, Lalitpur, NepalLulu is an indigenous breed of cattle (Bos taurus) found in high altitude regions of western Nepal. Population of Lulu cattle has been declining due to introgression with other exotic breeds to increase milk productivity. Here we aimed at finding potential approach for conserving Lulu cattle and its assets by studying the milk contents and investigating which variant of beta-casein protein is present in this breed. Beta caseins are an abundant protein in cow milk with A1 and A2 being the most common genetic variants of this protein. Consumption of A1 type of milk has numerous health-related complications whereas A2 type of milk has numerous human health promoting factors. We used restriction fragment length polymorphism (RFLP) for determining the A1 and A2 variant of beta casein in Lulu cattle. For performing DNA extraction, we collected (n = 18) blood samples of Lulu from Mustang and (n=17) Nepal Agriculture research council farm. The amplified fragments in 3% agarose at 251bp and 213bp respectively confirmed the presence of both A1 and A2 gene in Lulu; however, A2 was of greater abundance. Our study indicated that Lulu has A2 variant of beta-casein predominantly. The gene frequency of A1A1 is 0, A1A2 is 0.06 and A2A2 is 0.94. We further found that the allele frequency of A1 and A2 is 0.03 and 0.97 respectively. We designed special primer for sequencing CSN2 genes since A2 type beta casein gene was predominantly seen on Lulu. The sequencing result further supports our RFLP result as most of our samples have “C” nucleotide SNP in amplified CSN2 gene sequence. The Chi-square value of the current study is 0.04 which supports Hardy-Weinberg equilibrium inferring that Lulu cattle are still in the pure state, where there is no genetic introgression with the exotic breed for the sake of improvement of productivity.https://www.nepjol.info/index.php/JNARC/article/view/28115lulucsn2 genea2 variantrflpsequencing
spellingShingle Neena Amatya Gorkhali
Chhiring Sherpa
Nabina Budhathoki
Sudarshan GC
Srijana Lama
Prashanna Pokharel
Bhoj Raj Pokhrel
Saroj Sapkota
PCR Based Genotyping of Lulu Cattle of Nepal for A1, A2 Type Beta-caseins
Journal of Nepal Agricultural Research Council
lulu
csn2 gene
a2 variant
rflp
sequencing
title PCR Based Genotyping of Lulu Cattle of Nepal for A1, A2 Type Beta-caseins
title_full PCR Based Genotyping of Lulu Cattle of Nepal for A1, A2 Type Beta-caseins
title_fullStr PCR Based Genotyping of Lulu Cattle of Nepal for A1, A2 Type Beta-caseins
title_full_unstemmed PCR Based Genotyping of Lulu Cattle of Nepal for A1, A2 Type Beta-caseins
title_short PCR Based Genotyping of Lulu Cattle of Nepal for A1, A2 Type Beta-caseins
title_sort pcr based genotyping of lulu cattle of nepal for a1 a2 type beta caseins
topic lulu
csn2 gene
a2 variant
rflp
sequencing
url https://www.nepjol.info/index.php/JNARC/article/view/28115
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