Cloning of Yak SIRT1 Gene and Its Expression Pattern in Testis at Different Development Stages

doi:10.7668/hbnxb.20190855

Abstract

Abstract: This research was conducted to clone the SIRT1 gene in yak, predicated the structure and function of SIRT1 protein, identified its expression pattern in different tissues and testis of different ages. Nine healthy male yaks were divided into in calve (0.5 to 1 year), juvenile (2 to 3 years) and adult (4 to 5 years) groups (three in each group). Three samples from the same age were considered as biological duplications. The samples of liver, heart, spleen, brain, muscle, small intestine from adult sage and testis from three stages were collected. Total RNAs in different testis were extracted respectively and the sequence of yak SIRT1 gene was cloned by reverse transcription PCR(RT-PCR). Different bioinformatics softwares were used to predicate the homology of SIRT1 gene sequence, amino acid sequence, structure and properties of SIRT1 protein. Real-time PCR was applied to identify the mRNA expression profile of SIRT1 in different tissues of yak and chromogenic in situ hybridization(CISH) was applied to analyse the localization of SIRT1 mRNA in yak testis. Western Blot was conducted to detect the expression pattern of SIRT1 protein in yak testis of different stages. The open reading frame (ORF) of yak SIRT1 was 1 866 bp and encoded 621 amino acids. Sequence alignment analysis showed yak SIRT1 had a high sequence similarity with other mammals. Structure prediction indicated that SIRT1 was a hydrophilic liposoluble protein with a conserved silent information regulator 2 (Sir2) catalytic core domain. The mRNA of SIRT1 was widely expressed in different tissues of yak and was highly expressed in testis, ovary and liver. The testicular SIRT1 protein was highly expressed in calves and the expression decreased significantly with the growing of age. SIRT1 mRNA localized in multiple kinds of cells except spermatid. This research provides a theoretical basis for further understanding the mechanism of SIRT1 in regulating yak testicular development and is of great importance in providing new insights in improving yak reproductive performance in the future.

Key words: Yak, Histone deacetylation, SIRT1, Gene cloning, Testis

CLC Number:

Q78
S823

YIN Shi, QIN Wenchang, WANG Bin, YANG Liuqing, ZHOU Jingwen, LI Jian. Cloning of Yak SIRT1 Gene and Its Expression Pattern in Testis at Different Development Stages[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(4): 203-210. doi: 10.7668/hbnxb.20190855.

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