广灵驴CAPN1基因克隆、生物信息学分析及表达研究

doi:10.16431/j.cnki.1671-7236.2020.03.002

Abstract

Abstract: This study was aimed to clone the calcium protease 1 (CAPN1) gene CDS and analyze its bioinformatics,and identify its expression in different tissues of Guangling donkey.The CAPN1 gene was amplified,cloned and sequenced,the physicochemical,subcellular localization,modified structure and protein structure of CAPN1 protein were predicted using bioinformatic methods.The expression level of CAPN1 gene in heart,liver,spleen,lung,kidney and longissimus dorsi muscle of Guangling donkey were analyzed by Real-time PCR.The results showed that the length of CAPN1 gene CDS was 2 148 bp,and encoded 715 amino acids,which had been submitted to GenBank,the accession number was MN158194.The homology of nucleic acid sequence of CAPN1 gene in Guangling donkey was 99.7%,92.3%,92.5%,92.0%,92.5%,85.9% and 92.9% with Equus caballus,Ovis arise,Bos taurus,Homo sapiens,Capra hircus,Mus musculus and Sus scrofa,respectively.The molecular weight,isoelectric point,instability index of CAPN1 protein were 82.01 ku,5.59 and 36.42,respectively,and the average hydrophobic index was -0.374.CAPN1 protein had no signal peptide and no transmembrane region.The secondary structure of CAPN1 protein was consisted of random coil,alpha helix,beta turn and extended chain.The CAPN1 gene was expressed in all six tissues,the expression in longissimus dorsi muscle was the highest,followed by liver,and the expression in heart was the lowest.This experiment successfully cloned CAPN1 gene of Guangling donkey and analyzed its expression in different tissues,it provided a theoretical basis for further study of the expression and regulation function of CAPN1 gene in the muscle tenderness and developing the meat products industry of local species Guangling donkey.

Key words: Guangling donkey; CAPN1 gene; cloning; bioinformatics analysis; tissue expression

CLC Number:

S822

ZHAO Jingwei, SUN Yutong, LI Wufeng. Cloning,Bioinformatics and Expression Analysis of CAPN1 Gene in Guangling Donkey[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(3): 655-665.

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Cloning,Bioinformatics and Expression Analysis of CAPN1 Gene in Guangling Donkey

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