Cloning of Yak TNFAIP6 Gene and Its Temporal Expression in Ovarian Activity

doi:10.13560/j.cnki.biotech.bull.1985.2019-0596

Abstract

Abstract:

The aims of this study are to investigate the sequence characteristics of yak（Bos grunniens））TNFAIP6 gene and to analyze its temporal expression differences in different tissues and ovary at different stages of estrous cycle. Total RNA and protein from the heart,lung,spleen,kidney,liver,uterus,small intestine,gastric,muscle,and ovary tissues at different stages of estrous of healthy female yaks were extracted. The coding sequences of TNFAIP6 gene were cloned by RT-PCR and bioinformatics analysis of it was conducted. The expression level of TNFAIP6 gene in different tissues was detected by semi-quantitative PCR. Furthermore,the expression levels of protein and mRNA in yak ovary at different stages of estrous cycle were detected by Western blot and qRT-PCR. The collected data were statistically analyzed. The results showed that the CDS region of yak TNFAIP6 gene was 830 bp,encoding 279 amino acids. The analysis of proteins indicated that TNFAIP6 was hydrophilic and acidic-stable protein,had non-transmembrane domain and signal peptide,including Link and CUB structural domains. The secondary and tertiary structure of it was mainly composed of random coil and α-helix. Through homology alignment,the yak TNFAIP6 had high homology with cattle and wild yak. TNFAIP6 gene of yak from semi-quantitative PCR was widely expressed in various tissues and its expression in ovary,uterus,spleen and muscle was relatively higher than in other tissues. The expression level of TNFAIP6 in the follicular stage was significantly higher than that in the other two stages in ovary（P<0.01）,while there was no significant difference between the corpus luteum stage and red body stage（P>0.05）. The results of Western blot were basically consistent with qRT-PCR. In sum,the TNFAIP6 gene may be involved in the reproductive action of yak ovary,and the result provides basic reference in exploring the action mechanism of TNFAIP6 in the activity of yak ovary.

Key words: yak, ovary, TNFAIP6, temporal expression

MA Hong-cheng, XIONG Xian-rong, MU Song-yin, HAI Zhuo, QIN Wen-chang, LI Jian. Cloning of Yak TNFAIP6 Gene and Its Temporal Expression in Ovarian Activity[J]. Biotechnology Bulletin, 2019, 35(11): 46-54.

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