Multidimensional Characterization and Adaptive Differentiation of Leptin Gene in Eothenomys miletus: Based on Bioinformatics and System Evolution Analysis

Background: To investigate the differences between leptin gene in Eothenomys Miletus and other 55 mammal species, this study conducted a comparative analysis of multidimensional characteristics and adaptive differentiation of leptin genes. Methods: In this study, molecular biology experimental strategies have been used to extract and reverse transcription, gene cloning and sequencing of E. miletus and bioinformatics evaluation techniques have been used to analyze the sequence and structure, multi-sequence alignment and evolutionary evaluation of E. miletus and ecological and distribution statistical strategies had been used to analyze the leptin genes of 56 mammalian species such as E. miletus. Result: Leptin gene in E. miletus contained a Sec/SPI signal peptide, with α-helices dominating its structure. A model was constructed using leptin-LepR trimer (8 × 80.1. C leptin) as template. It encodes 127 amino acids with high proportions of leucine and serine; its molecular formula is C824H1353N221O252S8 (2658 atoms) with molecular weight of 18644.62 and theoretical isoelectric point of 5.86. The protein is unstable and hydrophilic, yet more hydrophobic than Tupaia chinensis and Homo sapiens. E. miletus was genetically close to Microtus, distant from Lagomorpha and clusters with temperate Rodentia. Its codon preference differed from Lagomorpha, with moderate GC content, GC3 content and ENC values. Motifs 1-5 were conserved in its leptin. Functional annotation shows regions related to signal peptide, leptin and extracellular domains. The ω value of ~0.5661 suggests evolutionary conservation. These results provide basis for exploring leptin gene evolution in E. miletus and mammals, offering perspective on mammalian adaptation to extreme environments and high-altitude protection.