策勒黑羊ERα基因克隆、生物信息学分析及卵巢组织表达研究

doi:10.16431/j.cnki.1671-7236.2022.05.009

Abstract

Abstract: 【Objective】 The aim of this experiment was to clone the estrogen receptor α (ERα) gene sequence in Cele Black sheep,and bioinformatics analysis was carried out.At the same time,the expression of ERα gene in ovary of Cele Black sheep were measured.【Method】 The ERα gene sequence of Cele Black sheep was obtained by RT-PCR and TA cloning,and bioinformatics analysis was carried out by online softwares.The expression of ERα gene in ovary of Cele Black sheep at follicular phase,luteal phase and 30 days of pregnancy were detected by Real-time quantitative PCR.【Result】 The length of ERα gene sequence in Cele Black sheep was 1 791 bp,and encoded 596 amino acids,with the highest similarity with Capra hircus and Bos taurus.Bioinformatics analysis results showed that the molecular formula of the protein was C₂₉₃₀H₄₆₀₀N₈₂₂O₈₆₂S₄₁,the theoretical isoelectric point (pI) was 7.32,the instability index was 50.33,which was an unstable hydrophilic protein.There were 28 O-glycosylation sites and 54 phosphorylation sites,but no N-glycosylation sites.ERα protein was mainly located in nucleus and there was no transmembrane structure.The secondary structure of ERα protein was mainly alpha helix and random coil.Real-time quantitative PCR showed that ERα gene in Cele Black sheep was expressed in ovary at follicular phase,luteal phase and 30 days of pregnancy,compared with follicular phase,the expression of ERα gene in Cele Black sheep was decreased extremely significantly at luteal phase (P<0.01),and increased extremely significantly at 30 days of pregnancy (P<0.01).【Conclusion】 The length of ERα gene sequence in Cele Black sheep was 1 791 bp,and encoded 596 amino acids,which was a hydrophilic protein.The secondary structure of ERα protein was mainly alpha helix and random coil.ERα gene in Cele Black sheep was expressed in ovaries at follicular phase,luteal phase and 30 days of pregnancy.The results provided a basis for further exploration of ERα gene which played a role in ovarian follicle development,corpus luteum formation and pregnancy of Cele Black sheep.

Key words: Cele Black sheep; ERα gene; cloning; expression

CLC Number:

S823

ZHANG Jihu, MI Renisa, WANG Zixuan, LI Lin, XING Feng, LIU Shudong. Cloning and Bioinformatics of ERα Gene and Its Expression of Ovary Tissue in Cele Black Sheep[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1679-1687.

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Cloning and Bioinformatics of ERα Gene and Its Expression of Ovary Tissue in Cele Black Sheep

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