摘要
旨在探讨ZP 3基因在海兰褐蛋鸡中的表达特性,为后期研究ZP3基因在蛋鸡卵泡发育中的作用提供理论支撑。本研究以产蛋期50周龄(50 W)海兰褐蛋鸡为试验对象,利用RACE技术克隆ZP 3基因全长,并对编码区进行生物信息学分析。选择健康、体重相近的海兰褐蛋鸡6只,分别取心、肝、肺、肾、卵巢、胸肌、腿肌和不同等级卵泡构建表达谱。卵巢颗粒细胞经不同浓度(PBS、5 ng·mL^(-1)、10 ng·mL^(-1)、20 ng·mL^(-1))促卵泡素(follicle-stimulating hormone,FSH)处理后,提取总RNA,采用实时荧光定量检测ZP 3基因在各个样品的表达水平。结果表明,鸡ZP 3基因全长1415 bp,其中编码区1341 bp,共编码446个氨基酸,位于10号染色体上,包含9个外显子;其亲缘关系与猪最远;跨膜结构预测表示,其含有一个跨膜结构域和一个信号肽区域;对其亲疏水性进行分析,预测该蛋白为弱的疏水性蛋白,蛋白质结构主要由无规则卷曲构成。组织表达谱显示,ZP 3基因在鸡的卵巢中相对表达量最高。各等级卵泡ZP3表达分析显示,随着卵泡发育,ZP 3基因在成熟卵泡(F1)颗粒细胞层表达量最高;在使用FSH处理等级颗粒细胞后,发现ZP3表达量显著上调(P<0.05),暗示ZP3基因在颗粒细胞中受到FSH激素调节。本试验对ZP 3基因的结构进行了分析并预测该基因存在跨膜结构和信号肽区域。基因表达谱分析结果显示,ZP 3基因在卵泡发育过程中表达量逐渐升高且主要在颗粒细胞中表达,可能受到FSH的调节作用,因此预测,ZP 3基因可能与海兰褐蛋鸡繁殖功能相关。
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 1415 bp containing 9 exons,in which the coding region was 1341 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 ZP 3 gene and predicted that it contained the transmembrane structure and signal peptide region.Expression profiles revealed that the expression level of ZP3 gene increased grad
作者
朱帅鹏
王东雪
刘聪
孙君卫
杜振伟
孙桂荣
李文婷
ZHU Shuaipeng;WANG Dongxue;LIU Cong;SUN Junwei;DU Zhenwei;SUN Guirong;LI Wenting(College of Animal Science and Technology,Henan Agricultural University,Zhengzhou 450046,China)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2021年第12期3366-3374,共9页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
中原学者康相涛科学家工作室(30601985)
国家自然科学基金(31902144)。
关键词
海兰褐蛋鸡
ZP
3基因
基因克隆
生物信息学
组织表达谱
Hailan brown laying hens
ZP3 gene
gene cloning
bioinformatics
tissue expression profile
