Molecular Characteristics of Bosgrunniens FAF1 Gene and Its Expression in Ovaries,Fallopian Tubes and Uterus at Different Stages

doi:10.13560/j.cnki.biotech.bull.1985.2019-0958

Abstract

Abstract: FAF1（fas-associated factor-1）binds to Fas protein in a variety of cells,mediating the initiation of apoptosis,and its gene mutation maylead to the death of embryos in the mitotic phase. The purpose of this study is to explore the molecular characteristics of Bosgrunniens FAF1 gene and its biological role in reproductive cycle. The FAF1 gene was cloned from ovaries,fallopian tubes and uterus of female B.grunniensat 3 different stages（luteal,follicular and the third month of pregnancy, the third month of pregnancy is hereinafter referred to as pregnancy）. The expression levels of genes and proteins were detected and located by real-time fluorescence quantitative PCR（qRT-PCR）,immunohistochemistry and Western blot. As results,the coding region（CDS）of B. grunniens FAF1 gene was successfully cloned in this study,its length was 1 953 bp（GenBank：MK416195）,encoding 650 amino acids. Genetic traits analysis showed that the gene was highly conservative,its encoded protein was non-transmembrane and soluble protein containing 5 protein binding sites,mainly distributed in the nucleus（52.2%）,mitochondria（26.1%）,the cytoplasm（13.0%）,golgi body（4.3%）,and cytoskeleton（4.3%）. The results of qRT-PCR demonstrated that the expression of FAF1 gene in the ovary was the highestat the follicular stage,followed by the pregnancy stage and the lowest attheluteal stage.The expressionin the oviductwas the highestat theluteal stage. The expression in the ovarywas the highest atthe follicular stage,the second atthe pregnancy stage,and the lowestat the luteal stage. The protein level showed that the relative expression of FAF1 protein in the fallopian tube and uterus during pregnancy was significantly higher than that atthe follicular and lutealstages. The expression of FAF1 protein in the ovary was the highest in the lutealstage,followed by the follicular stage,and the lowest in the pregnancystage. Immunohistochemistry（IHC）results revealed that there was no significant difference in the expression sites of FAF1 protein in the same tissue at different stages. The main expression sites in ovary were ovarian reproductive epithelium,granulosa cells,follicular membrane cells and corpus luteum cells（follicular phase）. The main expression sites in the fallopian tube were mucosal epithelial cells. The main expression sites in the uterus were endometrial cells and uterine glands. In conclusion,the expressions of FAF1 gene and protein in different reproductive cycles of the same reproductive organ were significantly different,which indicated that it was of great significance to the physiological regulation of B.grunniens reproduction.

Key words: FAF1, reproduction, bioinformatics analysis, express

WANG Jing-yu, WANG Meng, PAN Yang-yang, WANG Jing-lei, ZHANG Rui, MA Rui, HU Xue-quan, QIU Xiao-fei, CUI Yan, YU Si-jiu, XU Geng-quan. Molecular Characteristics of Bosgrunniens FAF1 Gene and Its Expression in Ovaries,Fallopian Tubes and Uterus at Different Stages[J]. Biotechnology Bulletin, 2020, 36(7): 80-89.

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