海兰褐蛋鸡ZP3基因结构及其在卵泡中表达规律分析

doi:10.11843/j.issn.0366-6964.2021.012.005

Abstract

Abstract: The study aimed to explore the expression characteristics of ZP3 gene in Hailan laying hens, which could lay the theoretical foundations for elucidating the function of ZP3 gene in follicle development in chicken. Chicken ZP3 gene was cloned with rapid amplification of cDNA ends (RACE) using the follicle tissue from 50-week-old (50 W) Hailan laying hens. The sequence of CDS region of ZP3 was analyzed by bioinformatics tools. The expression profile of ZP3 gene was constructed using heart, liver, lung, kidney, ovary, chest muscle, leg muscle tissues and follicles of different grades from 6 healthy Hailan brown laying hens with similar body weight. Total RNA was obtained from the ovarian granulosa cells with the treatment of follicle-stimulating hormone (FSH) in different concentrations (PBS, 5 ng·mL^-1,10 ng·mL^-1,20 ng·mL^-1). The expression levels of ZP3 gene in different tissues were detected by real-time PCR. The total length of chicken ZP3 gene was 1 415 bp containing 9 exons, in which the coding region was 1 341 bp, encoding 446 amino acids, located at chromosome 10. Its genetic relationship was the farthest from swine. The prediction of transmembrane structure indicated that it contained a transmembrane domain and a signal peptide; By analyzing its hydrophobicity, it was predicted that the protein was a weak hydrophobic protein, and the protein structure was mainly composed of random curl. According to the results of gene expression profile, the expression of ZP3 gene was the highest in chicken ovary. Besides, the expression of ZP3 gene was the highest in granulosa cells of mature follicles (F1) relative to follicles of other degrades. After treating granulosa cells of different grades with FSH, the expression level of ZP3 gene was significantly increased (P<0.05), suggesting that ZP3 gene was regulated by FSH in granulosa cells. This stuay analyzed the structure of ZP3 gene and predicted that it contained the transmembrane structure and signal peptide region. Expression profiles revealed that the expression level of ZP3 gene increased gradually during follicular develop ment and was mainly expressed in granulosa cells, which may be regulated by FSH. Therefore, it was predicted that ZP3 gene might be involved in reproductive function of Hailan brown laying hens.

Key words: Hailan brown laying hens, ZP3 gene, gene cloning, bioinformatics, tissue expression profile

CLC Number:

S831.2

ZHU Shuaipeng, WANG Dongxue, LIU Cong, SUN Junwei, DU Zhenwei, SUN Guirong, LI Wenting. ZP3 Gene Structure of Hailan Brown Laying Hens and Its Expression Regularity in Follicles[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(12): 3366-3374.

References 31

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ZP3 Gene Structure of Hailan Brown Laying Hens and Its Expression Regularity in Follicles

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