猪PFKL基因克隆及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2020.02.001

Abstract

Abstract: This study was aimed to explore the physicochemical properties and structural characteristics of porcine semen 6-phosphofructokinase (PFKL) gene,elucidate the physiological role and regulatory mechanism of glycolysis in spermatogenesis.PFKL gene was amplified by RT-PCR,the obtained gene fragment was inserted into pUC57 vector for cloning and sequencing,and its amino acids sequence,transmembrane structure,modification site,secondary structure,tertiary structure,antigen site and functional annotation were predicted and analyzed by bioinformatics methods.The results showed that the total length of PFKL gene was 2 349 bp,encoding 782 amino acids,the molecular weight of protein was 83.819 ku,the isoelectric point was 6.96,and the molecular formula was C₃₆₇₄H₅₈₉₇N₁₀₅₁O₁₁₀₉S₄₀.The homology of porcine PFKL gene was more than 80.0% compared with the reference sequences of Mus musculus,Iumenta anatis,Ovis aries,Bos taurus,Pongo abelii and Homo sapiens.The protein structure had no transmembrane structure and signal peptide region,and contained 2 N-glycosylation modification sites,14 O-glycosylation sites and 35 phosphorylation sites.The structure of PFKL protein was mainly composed of random coil and alpha helix,and its homology with muscle phosphofructokinase and platelet phosphofructokinase was 68.24% and 70.84%,respectively.At the same time,the 27 antigenic sites were different in strength,flexibility,uniform distribution and high surface plasticity,and there were binding sites of PKC and PKA specific protein kinase,which were hydrophilic proteins with more potential B cell epitopes.The results of GO functional annotation analysis showed that PFKL gene had carbohydrate kinase activity and phosphofructokinase activity,and could participate in hexoses metabolism and monosaccharides metabolism.In conclusion,this reaults laid a molecular biological foundation for improving sperm quality and the incidence of sperm-egg binding.

Key words: PFKL gene; glycolysis; bioinformatics analysis

CLC Number:

GE Chenling, LI Xun, WANG Xiaoye, LIANG Yingying, CHEN Zhiying, DU Qian, TANG Yinsheng, HUANG Mingguang, HU Chuanhuo. A Cloning and Bioinformatics Analysis of Porcine PFKL Gene[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(2): 315-325.

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A Cloning and Bioinformatics Analysis of Porcine PFKL Gene

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