Development of the Real-Time PCR method for detection of ss52050737 polymorphism of lhcgr gene in cattle

DOI

10.2991/ispc-19.2019.78How to use a DOI?

Keywords

LHCGR; ss52050737 polymorphism; cattle; real-time PCR; TaqMan

Abstract

Embryo transfer is one of the key technique in reproductive animal biotechnology. One of its crucial step is the selection of donor cows that are most sensitive to the hormonal stimulation of superovulation. One of the promising genetic markers of the reproductive status of cattle is the gene encoding the luteinizing hormone/choriogonadotropin receptor (LHCGR). One of the single nucleotide polymorphisms in the lhcgr gene of cattle − 1401G>T (ss52050737) – was shown to be associated with superovulation traits, the number of oocytes and the quality of embryos. To date, PCR-RFLP is one of the most common methods for detection of genes polymorphisms. But the main disadvantages of the PCR-RFLP method are long duration and laboriousness of the analysis. The real-time PCR method with allele-specific hydrolysis probes (TaqMan) is an effective alternative to PCR-RFLP analysis. The aim of this work was to develop the method for detection of ss52050737 polymorphism of lhcgr gene in cattle based on real-time PCR. In the developed method, two primers flaking the 113-bp fragment of lhcgr gene, and two allele-specific TaqMan probes were used. The identification of 1401G/T alleles (ss52050737 polymorphism) was based on the comparison of VIC and FAM fluorescence, respectively. The developed method was validated by PCR-RFLP analysis on 195 samples of cows of Russian Black Pied cattle. The developed method allows genotyping up to 480 animals (depending on the model of a thermal cycler) for 1 hour and can be used in selection of donor cows with superovulatory potential.

Copyright

© 2019, the Authors. Published by Atlantis Press.

Open Access

This is an open access article distributed under the CC BY-NC license (http://creativecommons.org/licenses/by-nc/4.0/).

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TY  - CONF
AU  - Svetlana Kovalchuk
AU  - Anna Arkhipova
AU  - Аrina Tagmazian
PY  - 2019/06
DA  - 2019/06
TI  - Development of the Real-Time PCR method for detection of ss52050737 polymorphism of lhcgr gene in cattle
BT  - Proceedings of the International Scientific and Practical Conference “Digital agriculture - development strategy” (ISPC 2019)
PB  - Atlantis Press
SP  - 346
EP  - 348
SN  - 1951-6851
UR  - https://doi.org/10.2991/ispc-19.2019.78
DO  - 10.2991/ispc-19.2019.78
ID  - Kovalchuk2019/06
ER  -