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王浆高产蜜蜂工蜂幼虫发育期肽质量指纹图谱的建立
李华玮1, 何金环1, 郑 鸣1
郑州牧业工程高等专科学校 生物工程系
摘要:
【目的】研究王浆高产蜜蜂(Apis mellifera L.)工蜂幼虫发育期蛋白质的组成及功能,以探明其发育机理,为阐明该蜂种王浆高产机理提供理论依据。【方法】采用双向电泳法对王浆高产蜜蜂2,4,6日龄的工蜂幼虫进行蛋白质组分析,获得图谱中蛋白质的种类、表达量、等电点和分子质量等信息,并通过质谱分析、数据库检索,鉴定部分蛋白。【结果】在2,4和6日龄的浆蜂工蜂幼虫中分别检测到262,418,194个蛋白,利用质谱技术鉴定的48个浆蜂工蜂幼虫发育期高丰度蛋白中有22个蛋白被鉴定出来,它们均属于蜜蜂数据库。与营养相关的蛋白有5个MRJP2、3个MRJP3和1个LSP2,与物质能量代谢相关的蛋白包括Arginine kinase、Aldehyde dehydrogenase、Enolase、Phosphoglycerate mutase、ATP synthase alpha subunit和ATP synthase,3个热激蛋白分别是HSP 60、HSC 70-3和HSP 8,4个与氨基酸、脂肪酸、幼虫生长和细胞周期相关的蛋白分别是Fatty acid binding protein、imaginal disc growth factor、lethal Band-7-prohibitin和Ornithine aminotransferase。【结论】王浆高产蜜蜂幼虫的发育过程有大量基因参与表达和调控,是一个复杂而动态有序的过程;在王浆高产蜜蜂幼虫发育期,与物质能量代谢及脂肪酸、氨基酸代谢相关的蛋白表达上调,说明幼虫在整个发育过程中需要大量能量,新陈代谢逐渐旺盛;持续表达的热激蛋白对减轻逆境胁迫引起的伤害起到一定作用,保证了王浆高产蜜蜂幼虫发育的正常进行。
关键词:  王浆高产蜜蜂  蛋白质组  工蜂幼虫
DOI:
分类号:
基金项目:
Establishment of peptide quality fingerprint for high royal jelly producing bees (Apis mellifera L.) during worker larval development
Abstract:
【Objective】The purpose was to investigate the protein composition of worker bee larvae of high royal jelly producing bees (Apis mellifera L.).【Method】Two-dimensional gel electrophoresis (2-DE) was employed to establish the proteomic map of the larvae of high royal jelly producing worker bees on 2,4 and 6 days,then the protein expression abundance,molecular weight and pI were obtained.Part of high abundance proteins were identified by analysis of matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF/MS) and database research.【Result】262,418,and 194 proteins were detected on day 2,day 4 and day 6 during worker larval development period,respectively.Out of 48 high abundance proteins selected for MS fingerprinting,22 were identified as representing annotated genes of the honey bee.These included five MRJP2,three MRJP3 and one LSP2 that were related to nutrient proteins,6 proteins named Arginine kinase,Aldehyde dehydrogenase,Enolase,Phosphoglycerate mutase,ATP synthase alpha subunit and ATP synthase that were associated with carbohydrate metabolism and energy production,HSP 60,HSC 70-3 and HSP 8 that were three heat shock proteins,and 4 other proteins called Fatty acid binding protein,imaginal disc growth factor,lethal Band-7-prohibitin and Ornithine aminotransferase relating to the metabolism of amino acids,fatty acid metabolism,larval growth and cell cycle,respectively.【Conclusion】The result indicated that the larvae development was a sequential and complex gene controlled process.The altered expression of the metabolism of carbohydrates and energy production and proteins which related to fatty acid metabolism confirmed that a lot of energy was needed during the larvae development and the metabolism was gradually accelerated.Heat shock proteins continuously expressed could protect cells when undergoing various types of environmental stress.
Key words:  high royal jelly producing bee  proteome  larvae