猪MORC2基因克隆、生物信息学分析及组织表达定位

doi:10.16431/j.cnki.1671-7236.2022.07.001

Abstract

Abstract: 【Objective】 The aim of this study was to clone porcine Microrchidia family CW-type zinc finger 2 (MORC2) gene,analyze the sequence characteristics using bioinformatics,and detect the expression of MORC2 gene in different tissues and the localization of MORC2 gene in ovary of pigs.【Method】 The complete CDS region of MORC2 gene was amplified and cloned with porcine ovary cDNA as template,and the similarity comparison and phylogenetic tree construction were carried out.The sequence of porcine MORC2 protein were predicted by bioinformatics software.The expression of MORC2 gene in different tissues of pigs were detected by Real-time quantitative PCR.The location of MORC2 protein in porcine ovary was detected by immunohistochemical method.【Result】 The length of MORC2 gene CDS was 3 102 bp,encoding 1 033 amino acids.The amino acid sequence of porcine MORC2 protein had 94.8%,94.6%,94.9%,91.9%,93.8%,93.9%,80.8% and 64.3% similarity with Human sapiens, Pan troglodytes, Macaca mulatta,Mus musculus,Bos taurus,Ovis aries,Gallus gallus and Danio rerio,respectively.Phylogenetic trees revealed that pig was most closely related to primates,ruminants and rodents were next,and the farthest related was Danio rerio.The molecular weight of porcine MORC2 protein was 117.44 ku,the isoelectric point was 8.16,the half-life was 30 h,and there were no transmembrane structure and signal peptides,it was an unstable hydrophilic protein.There were 174 phosphorylation modification sites and 81 glycosylation modification sites of MORC2 protein,it was mainly located in cell nuclear,less in cytoplasm and mitochondria.MORC2 protein contained the classical MORC protein family structure (GHKL-ATPase,zf-CW and CC domain).Tissue expression profile results showed that MORC2 gene was widely expressed in 11 tissues of pigs,the expression was the highest in liver,which was significant higher than that in other tissues (P＜0.05),and the expression were the lowest in heart,lung and muscle,which was significant lower than that in other tissues (P＜0.05).Immunohistochemical results showed that MORC2 protein was expressed in both granulosa cells and membrane cells of healthy pigs,and the expression was higher,but less in atretic granulosa cells and membrane cells.【Conclusion】 The CDS of porcine MORC2 gene was obtained,it was widely expressed in various tissues.MORC2 protein was expressed in both granulosa cells and membrane cells of healthy pigs.The results provided a theoretical basis for further research on the molecular mechanism of MORC2 protein regulating porcine ovarian development and follicular atresia.

Key words: pigs; MORC2 gene; cloning; expression; location

CLC Number:

QI Nannan, QIAN Yonghang, LUO Gang, SITU Xuqi, LIU Jiying. Cloning, Bioinformatics Analysis and Tissue Expression Location of Porcine MORC2 Gene[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2431-2441.

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Cloning, Bioinformatics Analysis and Tissue Expression Location of Porcine MORC2 Gene

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