猪SENP1基因克隆、生物信息学分析与表达研究

doi:10.16431/j.cnki.1671-7236.2021.12.006

Abstract

Abstract: The objective of this study was to clone and bioinformatics analyze the CDS of porcine sentrin-specific protease 1 (SENP1) gene, and investigate the expression pattern of SENP1 gene in PK15 cells infected with Japanese encephalitis virus (JEV).The specific primers were designed according to the predicted sequence of porcine SENP1 gene (accession No.:XM_013997974.2) published in GenBank.The sequence of SENP1 gene CDS was cloned and sequenced by RT-PCR and T/A cloning technology, and bioinformatics analysis was carried out.Real-time quantitative PCR was used to analyze the expression pattern of SENP1 gene in JEV-infected PK15 cells at different time points (0, 12, 24, 36 and 48 h).The results showed that the full length of SENP1 gene CDS was 2 034 bp, encoding a total of 677 amino acids.The sequence alignment and phylogenetic tree analysis results showed that the similarity of amino acid sequence of porcine SENP1 with Capra hircus, Canis lupus, Homo sapiens and Bos taurus was 94.8%, 94.2%, 93.9% and 93.8%, respectively.The results indicated that porcine SENP1 was highly conserved among these species.The genetic relationship of SENP1 between Sus scrofa and Canis lupus was close.The results of bioinformatics analysis showed that the relative molecular weight of SENP1 protein was 76 825.76, the isoelectric point was 8.53, the half-life in vitro was 30 h, the instability coefficient was 53.59, and the total average hydrophilic index was －0.622.The porcine SENP1 protein contained no signal peptide sequence and transmembrane domain.The ratio of alpha helix, random coil, extended chain and beta turn in secondary structure was 31.31%, 46.68%, 16.25% and 5.76%, respectively.The tertiary structure analysis showed that there were 8 alpha helix and 7 beta turn regions in porcine SENP1 protein.Real-time quantitative PCR results showed that the expression of SENP1 gene was up-regulated in JEV infected PK15 cells, and the expression of SENP1 gene at 48 h after infection was significantly higher than control group (P<0.05).This study provided reference materials for further study of porcine SENP1 gene function.

Key words: pigs; SENP1 gene; cloning; bioinformatics analysis; Real-time quantitative PCR

CLC Number:

DAI Zhenglie, ZHU Jingjing, CHEN Zhenyu, ZHOU Xiaolong, WANG Han, LI Xiangchen, ZHAO Ayong, YANG Songbai. Cloning, Bioinformatics Analysis and Expression of Porcine SENP1 Gene[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(12): 4382-4390.

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Cloning, Bioinformatics Analysis and Expression of Porcine SENP1 Gene

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