草鱼与3中脊椎动物MHCIα基因的克隆及其多态性比较-预防兽医学专业论文.docxVIP

万方数据 万方数据 Abstract Major histocompatibility complex (MHC) is a gene group of inmense polymorphism in vertebrate genomes, and MHC class I molecules are composed by α chain and β2m microglobulin. The α chain showed high polymorphism, including extracellular domain (α1, α2, α3), transmenbrane region (TM) and cytoplasmic (CY), in which the peptide binding region (PBR) of α1 and α2 domains is capable for combining antigen peptide and inducing specific immune response. The molecular structure and functional diversity is caused by polymorphism of MHC class I, and leading to differences of specific pathogen resistance. Therefore, the investigation of polymorphic characteristics of MHC class I α chain is help to reveal the molecular evolution and immune mechanism. In this study, the genomic structure and polymorphic characteristics of MHC class I α chain of low vertebrate Ctenopharyngodon idellus (Ctid) were investigated, and compared with other three vertebrates including mouse, cattle and chicken. Firstly, total blood RNA of three individuals was extracted, the Ctid-MHC I α gene was amplified by RT-PCR method, then cloned into pMD18–T vector and transformed into E.coli DH5α competent cells, thus the 20 positive clones of each individual were sequenced. Secondly, the molecular characteristics of polymorphic Ctid-MHC I α gene was analysed by bioinformatics software, and compared to other vertebrate including mouse, cattle and chicken. The results showed that 1027bp specific band was identified and 54 Ctid-MHC I α cDNA sequences were obtained. 1-3 MHC I α gene types were existed in each individual, 5 Ctid-MHC I α gene sequences were picked and uploaded to GeneBank database, then the accession number of Ctid-MHC I α gene were obtained (KF831073–KF831077). At the same time, MHC class I α chain accession number of mouse (KF831063–KF831067), cattle (KF831068–KF831072) and chicken (KF274027–KF274031) were obtained as well. Sequence analysis results showed that Ctid-MHC I α gene en