猪精子发生候选基因PYGO2的分离、表达和亚细胞定位研究

doi:10.11843/j.issn.0366-6964.2021.09.012

Abstract

Abstract: The purpose of this study was to isolate coding sequence of PYGO2 gene in pig, obtain its multi-tissue mRNA expression patterns, structural characteristics of protein and localization in cell, and construct the protein-protein interaction network. The coding sequence of PYGO2 was cloned from testis tissue of adult BMI using RT-PCR. The bioinformatics methods was performed to dissect its gene structure and protein functions, compare amino acid homology in mammals, construct phylogenetic tree and protein-protein interaction network. PYGO2 mRNA expression profiles in 15 tissues were detected using qPCR technology. The subcellular location of PYGO2 protein was conducted by constructing the expression vector of pEGFP-C1-PYGO2 and transfecting into ST cell. The results showed that the CDS sequence of PYGO2 was 1 221 bp, encoding 406 amino acids, and located on the chromosome 4 of pig genome. The accession numbers of gene and amino acid in GenBank were KY644518 and AVB77243.1, respectively. The secondary structure of PYGO2 protein contained mainly random coil with hydrophodic N and C terminal. The homology analysis of amino acid sequences implied that the similarities were more than 97% between pig and other mammals, suggesting high conservation of PYGO2 in evolution. The analysis of interaction network of proteins represented that pig PYGO2 had interactions with 9 proteins, and interacted most intensively with BCL9 protein. Further, multi-tissue qPCR results suggested that PYGO2 mRNA were extensively expressed in the 15 detected tissues and with higher expression in gonads. The result of subcellular location in transfected ST cells indicated that PYGO2 protein was predominantly localized in the nucleus. This study obtained the coding sequence of PYGO2 gene, protein structure and location, interaction network of protein, expression characteristics of mRNA in multiple tissues, which will provide a reference for further explanation of molecular mechanisms on pig PYGO2 during spermatogenesis.

Key words: pig, pygopus family PHD finger 2 (PYGO2), bioinformatics analysis, protein-protein interaction network, expression analysis, subcellular localization

CLC Number:

S828.3

ZHAO Xiao, ZHANG Xia, FAN Li, YANG Zhong, HUO Jinlong, ZENG Ribin, LIU Lixian, HUO Hailong. Isolation, Expression and Subcellular Localization of Spermatogenesis Candidate Gene PYGO2 in Pig[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(9): 2491-2499.

References 36

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Isolation, Expression and Subcellular Localization of Spermatogenesis Candidate Gene PYGO2 in Pig

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