牦牛SMPX基因CDS区克隆及组织表达分析

doi:10.16431/j.cnki.1671-7236.2019.05.003

Abstract

Abstract:

This study was aimed to clone the CDS sequence of small muscle protein X-link (SMPX) gene,analyze its potential structure and function,and explore the expression levels of SMPX gene in different tissues of Bos grunniens.The CDS sequence of SMPX gene was amplified and cloned by RT-PCR,the amino acid sequence homology of SMPX gene was analyzed,and the phylogenetic tree was constructed.The physical and chemical properties,secondary structure and tertiary structure of SMPX protein were analyzed by bioinformatics softwares.The expression patterns of SMPX gene in right ventricle,gluteus maximus,lung and brain of Bos grunniens were detected by Real-time PCR.The results showed that the length of SMPX gene CDS and ORF sequences were 515 and 261 bp,respectively,which encoded 86 amino acids.The amino acid sequence homology of SMPX gene in Bos grunniens with Bos mutus,Bubalus bubalis,Canis lupus familiaris,Homo sapiens,Odocoileus virginianus texanus,Ovis aries,Pan troglodytes,Pantholops hodgsonii and Sus scrofa were 100%,97.7%,96.5%,96.5%,95.3%,95.3%,96.5%,94.2% and 91.9%,respectively,indicating that it was highly conserved in different species.Bioinformatics analysis represented that SMPX protein was unstable and hydrophilic.The secondary structure of SMPX mainly contained random coils and alpha helix.The SMPX protein had no signal peptide and transmembrane protein, and belonged to intramembrane protein.There were four phosphorylation sites in SMPX protein sequence.The results of subcellular localization showed that SMPX protein was located in the nucleus (52.2%),mitochondria (43.5%) and cytoplasm (4.3%).Real-time PCR results exhibited that the expression level of SMPX gene in right ventricle was significantly higher than gluteus maximus,lung and brain (P<0.05).This study might provide reference data for further study of physiological functions and regulatory mechanisms of SMPX gene in Bos grunniens.

Key words: Bos grunniens; SMPX gene; cloning; bioinformatics; organizational expression

CLC Number:

GUO Liang, WANG Jikun, WANG Hui, CHAI Zhixin, XIN Jinwei, JI Qiumei, ZHONG Jincheng. CDS Cloning and Tissue Expression Analysis of SMPX Gene in Bos grunniens[J]. , 2019, 46(5): 1273-1282.

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CDS Cloning and Tissue Expression Analysis of SMPX Gene in Bos grunniens

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