关键词: 水牛; 甲状旁腺素; 基因克隆; 生物信息学

Abstract:

The buffalo parathyroid hormone (PTH) gene was successfully cloned and analyzed by bioinformatics method referring to PTH gene of Bos taurus in this study,and the coding sequences (CDS) was 348 bp that encoded 115 amino acids.The CDS of the buffalo PTH gene showed 99%,93%,91%,97%,96% and 92% identity with Bos taurus, Sus scrofa, Equus caballus, Capra hircus, Ovis aries and Camelus ferus,respectively,indicating that the PTH gene was conservative in the course of evolution. The analysis of the phylogenetic tree showed that the nearest relationship existed between buffalo and Bos taurus. Three base mutations were found,two of them were synonymous mutation,one was missense mutation.Amino acid sequence analysis showed that the PTH protein of buffalo was alkaline,hydrophilic and stable whose formula was C₅₇₀ H₉₄₁ N₁₆₇ O₁₆₄ S₈,and the molecular weight was 13.01 ku.The secondary structure of PTH protein was mainly α-helices and random coil,consistent with the results of tertiary structure prediction. The results of subcellular localization showed that the PTH protein was in the cytoplasm(26.1%),mitochondria (21.7%),extracellular (including cell wall)(17.4%),endoplasmic reticulum (13.0%),nucleus (8.7%),golgi apparatus (4.3%) and others (8.7%),and the PTH might play a role of signal transducer or transcription regulation in the transport and binding,as well as cell envelope.

Key words: buffalo; parathyroid hormone (PTH); gene cloning; bioinformatics