牦牛跨膜附睾蛋白1基因的克隆及其在卵丘卵母细胞复合体中的表达分析

doi:10.16431/j.cnki.1671-7236.2021.04.015

Abstract

Abstract: The aim of this study was to analyze the molecular characteristics and the expression level of transmembrane epididymal protein 1 (TEDDM1) gene during follicle development of the yak.In this study,cumulus-oocyte complexes (COCs) were collected from small follicles (φ≤3 mm),medium follicles (φ=5-7 mm) and large follicles (φ≥8 mm) of yak during estrus,and total RNA was extracted and reverse transcription was conducted.The complete sequence of the CDS region of the TEDDM1 gene was sequenced.Bioinformatics soft-wares were used to analyze the molecular characteristics of CDS region sequence structure.Using GAPDH as reference gene,Real-time quantitative PCR was performed to reveal expression level of TEDDM1 in small,medium and large follicles.The results showed that the CDS region of yak TEDDM1 gene was 903 bp,encoding 300 amino acid,which was highly similar to Bos mutus,Bos taurus,Bison,Ovis and Bubalus bubalis,and was conserved in evolution.Yak TEDDM1 protein was a non-secretory unstable hydrophobic protein with seven helix structures,and the whole peptide chain crosses the cell membrane seven times,which was a typical G protein-coupled receptor.Twenty-one potential phosphorylation sites were distributed in the protein,including 1 tyrosine,3 threonine and 17 serine phosphorylation sites,only one N-glycosylation site and no O-glycosylation prediction site.Functional domain analysis showed that unknown functional domain protein family 716 domain existed in TEDDM1 protein,covering multiple transmembrane regions.Result of Real-time quantitative PCR analysis showed that the expression level of TEDDM1 in medium follicles was significantly higher than that in large follicles and small follicles (P<0.05),but there was no significant difference between large follicles and small follicles (P>0.05).The study provided the sequence of yak TEDDM1 gene and its expression characteristics in yak COCs,laid a theoretical foundation for further revealing the regulatory role of the yak TEDDM1 gene in follicular development.

Key words: yak; follicles; transmembrane epididymal protein 1(TEDDM1) gene; cloning; Real-time quantitative PCR

CLC Number:

S823

LI Heng, ZI Xiangdong. Cloning and Expression in Cumulus-oocyte Complexes of Yak TEDDM1 Gene[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(4): 1284-1293.

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Cloning and Expression in Cumulus-oocyte Complexes of Yak TEDDM1 Gene

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