Cloning and Bioinformatics Analysis of MAPK13 Gene in Lanzhou Fat-tailed Sheep

Abstract

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.

Key words: Lanzhou fat-tailed sheep, MAPK13 gene, RACE, Bioinformatics analysis

Jin Fangyuan, Xu Hongwei, Da Xiaoqiang, Zang Rongxin, Bai Jialin, Gao Xin, Cai Yong, Feng Yulan, Yang Jutian. Cloning and Bioinformatics Analysis of MAPK13 Gene in Lanzhou Fat-tailed Sheep[J]. Biotechnology Bulletin, 2014, 0(8): 94-101.

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