生物技术通报Issue(8)：94-101,8.
兰州大尾羊MAPK13基因cDNA克隆及生物信息学分析
Cloning and Bioinformatics Analysis of MAPK13 Gene in Lanzhou Fat-tailed Sheep
摘要
Abstract
The objective of this study is to reveal the biological function of mitogen-activated protein kinases(MAPK)gene in sheep and provide theoretical data for application by cloning MAPK and analyzing its sequence as well as genetic features. The specific primers on the CDS template of MAPK13 gene were designed and then the MAPK13 gene sequence of Lanzhou fat-tail sheep was cloned using RACE and RT-PCR technology. After that, its genetic characteristics were analyzed by bioinformatics methods. The 1 397 bp full-length cDNA of MAPK13 of Lanzhou fat-tailed sheep was obtained, in which the length of CDS is 1 102 bp. It encodes 367 amino acids in total. It was predicted that the molecular weight of MAPK13 protein of Lanzhou fat-tail sheep is 42.29 kD and its theoretical isoelectric point is 8.82. It is a non-transmembrane hydrophobin with no signal peptide, which locates mostly in cytoplasm by the result of subcellular level prediction and does not belong to the secretory protein. Its amino acid sequence contains 19 phosphorylation sites, 3 glycosylation sites, 3 phosphorylation domains, 4 other domains and 1 LCR domain. Also, its secondary structure is mainly randomly curly. Nucleotide sequence analysis revealed that the gene nucleotide sequence of Lanzhou fat-tail sheep has some different from the known sheep MAPK13 mRNA(Genbank No.:NM_001139455.1). Base transition occur in the 852th site is(C vs. A), respectively, the corresponding amino acids in the 265th is(S vs. T). Base transition occur in the 951th site is(T vs. G), but the corresponding amino acids do not change.The phylogenetic tree indicates that Lanzhou fat-tail sheep is close to bos. The fact that the structure of MAPK13 of Lanzhou fat-tail sheep highly similar with other species indicates that the gene is highly conservative. The S_TKc domain in the sequence can transfer the gamma phosphoryl of ATP to the serine/threonine residues of protein, leading to function imbalance and expression change in a series of obesity related gene.关键词
兰州大尾羊/MAPK13基因/cDNA/末端快速扩增/生物信息学Key words
Lanzhou fat-tailed sheep/MAPK13 gene/RACE/Bioinformatics analysis引用本文复制引用
金方园,徐红伟,达小强,臧荣鑫,柏家林,曹忻,蔡勇,冯玉兰,杨具田..兰州大尾羊MAPK13基因cDNA克隆及生物信息学分析[J].生物技术通报,2014,(8):94-101,8.基金项目
国家自然科学基金项目（31160440，31260533，31360529）,甘肃省科技支撑计划项目（1011NKCA051）,兰州市科技计划项目（2011-1-113），西北民族大学研究生科研创新项目 （）
