
意大利蜜蜂amPGAM2基因的克隆、 序列特征及表达分析
Cloning, sequence characterization and expression of the Apis mellifera amPGAM2 gene
欧阳霞辉;彭 帅;徐文凯;朱雪蕾;郑相相;陈 红
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DOI:10.7679/j.issn.2095-1353.2020.091
作者单位:西北民族大学生命科学与工程学院;西北民族大学生命科学与工程学院;西北民族大学生命科学与工程学院;西北民族大学生命科学与工程学院;西北民族大学生命科学与工程学院;西北民族大学生命科学与工程学院
中文关键词:意大利蜜蜂;PGAM2基因;糖酵解;序列特征;荧光定量
英文关键词:Apis mellifera; PGAM2 gene; glycolysis; sequence characteristics; fluorescence quantification
中文摘要:
【目的】 磷酸甘油酸变位酶(PGAM)是糖酵解和葡萄糖异生途径中一种发挥重要作用的蛋白酶,本研究拟明确amPGAM2基因的序列特征及表达模式。【方法】 以意大利蜜蜂Apis mellifera为研究对象,克隆了amPGAM2基因的cDNA序列,分析了其序列特征及其在工蜂、雄蜂、蜂王的不同发育时期的表达模式。【结果】 序列特征分析表明,克隆所得序列全长976 bp,包含1个完整的开放阅读框,共编码254个氨基酸。该基因核苷酸序列与中华蜜蜂Apis cerana(98.4%)高度相似,并存在15个潜在抗原表位、9个磷酸化位点和5个组氨酸磷酸酶域活性部位,属于组氨酸磷酸酶超家族,是一种二磷酸甘油酸依赖性的可溶中性稳定蛋白。荧光定量PCR结果表明,amPGAM2基因在不同品级、不同发育时期的表达差异显著(P<0.05)。工蜂卵期3日龄、幼虫期5日龄表达最高,雄蜂成虫期表达最高,蜂王幼虫期4日龄表达最高,且工蜂、雄蜂及蜂王由卵孵化成幼虫阶段和由红眼蛹羽化至成虫阶段,其表达都呈上升趋势。【结论】 本研究结果推测amPGAM2基因在卵的孵化及精卵发生过程中具有重要作用,为进一步认识意大利蜜蜂生殖发育过程中的能量代谢提供理论基础。
英文摘要:
[Objectives] Phosphoglycerate mutase (PGAM) plays an
important role in the glycolysis and gluconeogenesis of protease. This study
intends to clarify the sequence characteristics and expression patterns of the amPGAM2 gene. [Methods] The cDNA sequence of the Apis mellifera amPGAM2 gene was cloned, and its sequence characteristics and expression in different
castes and developmental stages; workers, drones and queens, analyzed. [Results] Sequence analysis indicates that the full
length of the cloned sequence is 976 bp, and contains a complete open reading
frame encoding 254 amino acids. The nucleotide sequence of the gene is highly
homologous to that of Apis cerana (98.4%). The gene has 15 potential
antigen epitopes, 9 phosphorylation sites and 5 histidine phosphatase domain
active sites and encodes a glycerol diphosphate-dependent soluble neutral
stable protein that belongs to the histidine phosphatase superfamily. The
results of RT-qPCR indicate that expression of amPGAM2 is significantly
different in different castes and developmental stages. Expression in workers
was highest at 3 days of age in the egg stage and at 5 days in the larval
stage. The highest expression in drones and queesn was in the adult stage and
in 4 day-old larvae, respectively. Expression in workers, drones and queens
increased from hatching to the larval stage, and from the red-eye pupal
eclosion to the adult stage. [Conclusion] The results suggest that the amPGAM2 gene plays an important role in
hatching and spermatogenesis. This paper provides a theoretical basis for
improving understanding of the energy metabolism during reproductive
development in Apis mellifera.