Abstract:

In order to explore the structure and biological function of Ras related nuclear protein gene(RAN)in yak and resolve how it regulates the proliferation of yak cells and participates in protein synthesis,the CDS region sequence of RAN gene of Sangsang yak was cloned by RT-PCR using Sangsang yak kidney tissue cDNA as template,and bioinformatics analysis was carried out by a variety of software and online tools,the expression of the RAN gene in seven tissues of Sangsang yaks was detected by qPCR technology.The results showed that the CDS region of RAN gene in Sangsang yak was 651 bp in length,encoding 216 amino acids.Through homology comparison,it was found that the genetic relationship between Sangsang yak and wild yak and zebu was the closest,with a similarity of 99.2%,and the farthest from chicken,reaching 86.6%.The prediction results of RAN protein analysis showed that the molecular weight of the protein was 24.423 11 ku,the theoretical isoelectric point was 7.01,the total number of atoms was 3 449,and the molecular composition was C₁₁₀₉H₁₇₂₅N₂₉₅O₃₁₃S₇.RAN protein had no transmembrane structure and no potential sites for N-glycosylation,with 36 phosphorylation sites.The affinity and hydrophobicity were predicted and the instability coefficient was calculated,and it was found that the protein was a stable hydrophilic protein.According to the subcellular localization,the protein was found to be present in the Golgi apparatus,mitochondria,nucleus and cytoplasm in yak cells.Predictions of the RAN protein structure revealed that its higher-order structure consisted mainly of α-helices and did not contain β-turns.Protein interaction network results showed that there was an interaction between the RAN protein and RAN Binding Protein 1(RANBP1),RAN Binding Protein 2(RANBP2),RANGTPase activating protein 1(RANGAP1)and other proteins of Sangsang yak,and there was also an interaction between them.The expression level of the RAN gene in yak testicular tissue was significantly higher than in other tissues,while no expression was detected in muscle tissue.It successfully cloned the CDS region of the RAN gene and completed its bioinformatics analysis,and the expression of this gene in the tissue of Sangsang yak was also studied,and it was found that it played an important role in the development of the reproductive system,cell proliferation,disease prevention and control,and participation in protein synthesis.

Key words: Sangsang yak, RAN gene, Gene cloning, Bioinformatics analysis, Expression

CLC Number: 

• S823.85

• Q78基因工程(遗传工程)