Sequence analysis of AK1 gene from Jingyuan chicken and construction of its eukaryotic expression vector

Objective  To explore the structure and function of protein encoded by AK1 gene screened by transcriptome sequencing of Ningxia local breed Jingyuan chicken, and construct its eukaryotic expression vector.

Method  According to the original chicken AK1 gene sequence published on GenBank, specific primers were designed for its CDS region, and the CDS region SNPs of AK1 gene were screened quickly by cloning and sequencing. The eukaryotic expression vector of AK1 gene was constructed. The bioinformatics function of the coding region was analyzed.

Result  The full length of AK1 gene coding region was 585 bp, which encoded 194 amino acids. AK1 protein had no transmembrane domain, and it had two CpG islands, 18 phosphorylation sites and 10 antigenic epitopes. AK1 protein was a stable water-soluble protein, and the spatial structure was mainly α-helix and irregular crimp. The results of subcellular localization showed that AK1 protein was mainly located in the cytoplasm, and GO enrichment analysis showed that AK1 gene was also enriched in the cytoplasm, which was consistent with the result of subcellular localization. Gene co-expression analysis of genes interacting with AK1 showed that AMPD1 and PKM2 coexpressed with AK1 gene, and the co-expression coefficients were 0.116 and 0.063, respectively. The AK1-pEGFP-N1 vector was successfully constructed.

Conclusion  The results of this study provide a scientific basis for further study of the function of AK1 protein and AK1 as an inosinic acid-related gene.