Isolation, Sequence Molecular Characteristics and Tissue Expression of MAPK10 Gene in Lijiang Pig

PurposeTo isolate mitogen-activated protein kinase (MAPK10) gene from Lijiang pig and to analyse the expression of mRNA and protein in tissue.

MethodsThe MAPK10 gene in Lijiang pig was amplified by PCR, and the bioinformatics software was used for sequence analysis. The relative expression level of MAPK10 gene in nine tissues of Lijiang pig including brain, liver, heart, spleen, lung, kidney, pancreas, back fat and longissimus dorsi, which was detected by real-time fluorescence quantitative PCR. Taking the Large White pig as a control, real-time fluorescent quantitative PCR and Western-blotting were used to detect the difference in mRNA and protein expression in the back fat of Lijiang pig and Large White pig.

ResultsThe total length of MAPK10 gene sequence was 1281 bp, which encodes 426 amino acids. MAPK10 protein was a hydrophilic protein without transmembrane domain and signal peptide. There were 44 phosphorylation sites and two glycosylation sites. The secondary structure of Lijiang pig MAPK10 was mainly consisted of α-helices and random coils. The homology between the sequence of Lijiang pig MAPK10 gene and that of human, cattle, goat, sheep, mouse, dog, monkey, and chicken was 95.3%, 93.5%, 93.2%, 93.0%, 92.6%, 91.2%, 90.5% and 85.6%, respectively. MAPK10 mRNA was widely expressed in the nine tissues of Lijiang pigs with highest expression level in brain. The mRNA expression level of MAPK10 gene in back fat of Lijiang pigs was extremely significantly lower than that of the Large White pig (P<0.01), and the protein expression level was also lower than that of the Large White pigs (P>0.05).

ConclusionMAPK10 gene can be used as a candidate gene for studying fat deposition traits in Lijiang pigs, and the result can provide reference for the study of fat deposition of pigs.