树鼩LCAT全长编码序列的克隆与分子特征分析OA

Objective To obtain the full-length coding sequence of the Chinese tree shrew lecithin cholesterol acyltransferase(tsLCAT)gene and carry out its molecular characterization.Methods Total RNA was extracted from Chinese tree shrew liver tissue,and the full-length cDNA of tsLCAT was cloned by rapid Amplification of cDNA ends technology.A phylogenetic tree was constructed for evolutionary relationship analysis using MEGA software.The molecular characteristics and biological functions were predicted using ExPASy-ProtParam,SignaIP,and MusiteDeep,and protein structure prediction and alignment were achieved using AlphaFold3.The mRNA and protein expression levels of tsLCAT in 11 tissues,including liver,pancreas,and kidney,were detected by reverse transcription-quantitative polymerase chain reaction and Western blot,respectively.Results Two transcript variants of tsLCAT were obtained:tsLCAT X1(full length 1 384 bp,encoding 440 amino acids)and tsLCAT X2(full length 1 400 bp,encoding 438 amino acids),with tsLCAT X1 being the major transcript.Phylogenetic analysis revealed that tsLCAT had a closer evolutionary relationship with primates than with rodents(such as mice and rats),and tsLCAT showed the highest amino acid sequence identity with human and chimpanzee LCAT.Tissue expression profiling indicated that tsLCAT mRNA expression levels were highest in the liver,and protein levels were enriched in the liver and pancreas.Conclusions The full-length coding sequence of tsLCAT was successfully obtained and characterized.Systematic analysis of its molecular characteristics,protein structure,and biological functions provides key molecular evidence for the use of Chinese tree shrews as a research model for LCAT-related diseases.