Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (6): 279-290.doi: 10.13560/j.cnki.biotech.bull.1985.2021-1152

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Cloning of Bos grunniens TGF-β1 Gene and Its Expression in Major Organs of Female Reproductive System

WANG Nan(), ZHANG Rui, PAN Yang-yang, HE Hong-hong, WANG Jing-lei, CUI Yan, YU Si-jiu()   

  1. College of Veterinary Medicine/Technology and Research Center of Gansu Province for Embryonic Engineering of Bovine and Sheep & Goat,Gansu Agricultural University,Lanzhou 730070
  • Received:2021-09-07 Online:2022-06-26 Published:2022-07-11
  • Contact: YU Si-jiu E-mail:594302635@qq.com;sjyu@163.com

Abstract:

Transforming growth factor β1(TGF-β1)is a multifunctional growth and differentiation factor,which regulates many physiological and pathological processes of the body,and has important biological functions and broad application prospects. In this study,we collected ovarian,oviductal and uterine tissues from yaks during estrus and pregnancy,cloned the yak TGF-β1 gene,and performed the quantitative analysis of yak TGF-β1 at the gene and protein levels using quantitative real-time fluorescence PCR(qRT-PCR),immunohistochemistry(IHC),and protein immunoblotting(Western blot). The results showed that TGF-β1 gene(GenBank No.MZ004937)was highly conserved,and the nucleotide sequence differed from the TGF-β1 nucleotide sequence of Bos taurus,and the encoded amino acid was mutated from alanine to glycine. It was most closely related to Bos taurus,Bos indicus×Bos taurus,and most distantly related to Canine. The encoded protein was a stably hydrophilic transmembrane protein. TGF-β1 was expressed in ovary,oviduct and uterus tissues of yak during follicular phase,luteal phase and gestation phase. In ovary,the expression of TGF-β1 during gestation phase was significantly higher than that during follicular phase and luteal phase(P<0.05). In oviduct,the expression of TGF-β1 gene in gestation phase was significantly higher than that in follicular and luteal phases(P<0.01). In utero,the expression in follicular phase and gestation phase was significantly higher than that in luteal phase(P<0.05). Immunohistochemistry(IHC)results indicated that TGF-β1 was mainly expressed in ovarian reproductive epithelium,follicular membrane,granular layer of follicle,luteal cells,tubal mucosal epithelial cells,uterine gland(UG)and endometrium cells. The results will provide basic data for further exploring the molecular mechanism of TGF-β1 in reproductive physiology of yaks,and provide theoretical basis for exploring the adaptability of plateau mammals to alpine environment.

Key words: yak, TGF-β1, gene cloning, bioinformatics analysis, expression