猪ASB2基因CDS区克隆、生物信息学分析及表达规律研究

doi:10.16431/j.cnki.1671-7236.2020.05.001

Abstract

Abstract: The objective of this study was to clone the complete CDS region of porcine ankyrin repeat and suppressor of cytokine signalling box containing protein 2 (ASB2) gene,analyze the CDS sequence and the basic characteristics of proteins by bioinformatics methods,and explore its expression profiles in different tissues of one-day-old Jinfen White pig and satellite cell myoblast induction.Based on the predicted nucleotide sequence of pig ASB2 gene in GenBank.Primers were designed to amplify the CDS region of ASB2 gene by RT-PCR using cDNA of longissimus dorsi muscle as template.The amino acid sequence and the structure and function of ASB2 were analyzed using bioinformatics software.The expression profiles in different tissues and during the procession of myogenic differentiation of satellite cells were investigated by Real-time quantitative PCR.The results showed that the complete CDS region of porcine ASB2 gene was 1 824 bp in length,which encoded a total of 607 amino acids.The nucleotide sequence of porcine ASB2 gene had the highest similarity with those of goat and cow.Bioinformatics analysis results showed that ASB2 was a hydrophilic protein,including 11 O-glycosylation sites,1 N-glycosylation site,and no signal peptide.There were 11 ANK motifs and 1 SOCS motif.The secondary structure of the protein was predicted to be 43.99%,40.36%,10.05% and 5.60% with random coil,alpha helix,beta turn and extended strand.Real-time quantitative PCR results showed that the highest expression level was found in porcine psoas muscle,and followed by longissimus dorsal muscle and heart,which were extremely significantly different from those in other tissues (P<0.01).During the procession of myogenic differentiation of satellite cells,the expression of ASB2 gene was increased at the beginning,reached the peak after 2 d induction,then decreased,which suggested that ASB2 gene might be involved in regulating muscle growth.The results of this experiment provided a reference for further exploring the function and mechanism of porcine ASB2 gene.

Key words: pig; ASB2 gene; cloning; sequence analysis; expression

CLC Number:

QIN Benyuan, YANG Shuai, ZHANG Yanwei, LI Wenxin, LIU Hong, WU Yiqi, ZHANG Xuelian, LI Wenxia, YANG Yang, CAI Chunbo, GAO Pengfei, GUO Xiaohong, LI Bugao, CAO Guoqing. Study on Cloning,Bioinformatics Analysis and Expression Profile of ASB2 Gene CDS in Pig[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(5): 1289-1298.

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Study on Cloning,Bioinformatics Analysis and Expression Profile of ASB2 Gene CDS in Pig

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