Abstract:

In order to explore the sequence characteristics of yak Akirin1 gene and understand its tissue expression characteristics in detail.Gannan yak muscle tissue cDNA was used as a template to clone the CDS sequence of Gannan yak Akirin1 gene by RT-PCR technology,and bioinformatics analysis was performed using a variety of biological online software and tools.The qPCR technique was used to detect the difference expression of Akirin1 gene in Gannan yak heart,liver,spleen,lung,kidney,muscle,fat and testis tissues.The results showed that the CDS of Akirin1 gene in Gannan yak was 579 bp,encoding 192 amino acids.The protein molecular formula encoded by the gene was C₉₆₄H₁₅₂₅N₂₇₅O₂₉₁S₈,the theoretical isoelectric point of the protein was 8.91,the instability coefficient and the total average hydrophilicity were 84.14 and -0.822,respectively,which belonged to the unstable hydrophilic protein.There were 29 potential phosphorylation sites,including 14 serines,11 threonines and 4 tyrosines.There was no signal peptide and transmembrane helix structure.Subcellular localization showed that it mainly existed in the nucleus and belonged to non-secretory protein.The high-level structure of the protein was mainly composed of 39.06% α-helix and 56.25% random coil structure.Homology analysis showed that the genetic relationship between yak and cattle was the closest,which was in line with the actual situation,indicating that the Akirin1 gene was relatively conservative in the evolution process.Real-time Fluorescence Quantitative results showed that Akirin1 gene was expressed in heart,liver,spleen,lung,kidney,muscle and adipose tissue of adult Gannan yak,and the relative expression in kidney tissue was the highest,which was significantly higher than that in other tissues,and the expression in muscle and liver tissue was relatively low.This experiment provides a theoretical basis for further study on the functional characteristics of yak Akirin1 gene.

Key words: Yak, Akirin1 gene, Cloning, Bioinformatic analysis, Tissue expression nalysis