Purposes Aspartate aminotransferase, cytoplasmic, also called glutamic-oxaloacetic transaminase 1, is an enzyme that is encoded by the nuclear GOT1 gene. It catalyzes the reversible transamination reaction from L-aspartate and α-ketoglutarate to L-glutamate and oxaloacetate, and plays a crucial role in the metabolic regulation. The aim of this study was to amplify GOT1 gene from Banna mini-pig inbred line (BMI) and to obtain its expression pattern.
Methods The GOT1 gene was cloned using RT-PCR method from BMI. Nucleotide and protein sequences of GOT1 were utilized to carry out bioinformatics analysis. The semi-quantitive RT-PCR technology was applied to analysis the mRNA expression profiles of tissues.
Results A cDNA of 1 484 bp (GenBank accession No. KU705636.1) of BMI GOT1 was obtained, which contained 1 242 bp opening reading frame. Bioinformatics analysis showed GOT1 gene localized to Sus scrofa chromosomes 14 and consisted of nine exons and eight introns without signal peptide sequences and transmembrane helix. GOT1 protein contained 36 potential phosphorylation sites and 1 O-GlcNAc glycosylation site and to be located in the cytoplasm with 94.1% certainty. The predicted amino acid sequence of the gene has high homology with the water buffalo (95.6%), cattle (95.4%), chimpanzee (93.0%), human (92.7%), cynomolgus monkey (92.5%), rat (90.3%) and mouse (90.1%). The secondary structure of BMI GOT1 was mainly consisted of α-helices, random coils and and extended strands were medium; whereas beta turns were locally present. Its tertiary structure was also constructed by homology modeling. GOT1 mRNA was widely expressed in the 15 tissues of BMI with highest expression level in muscle.
Conclusion The above results will lay a foundation for further study of the gene about its physiological function and regulatory mechanisms in BMI.
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