摘要
通过比较家蚕微孢子虫(Nosema bombycis)与桑尺蠖来源微孢子虫之间的蛋白质组差异,探讨二者对家蚕的侵染力差异产生的原因。采用Percoll法纯化微孢子虫,再以玻璃珠破碎法提取2种来源的微孢子虫总蛋白质,经Bradford法测定蛋白质浓度后进行双向电泳(2-DE)。通过Imagemaster软件分析比较家蚕微孢子虫与桑尺蠖来源微孢子虫总蛋白质的2-DE图谱,可较为明显地发现后者缺少11个蛋白点,前者缺少2个蛋白点。对13个差异蛋白点进行质谱鉴定,有一个大小约40 kD的差异蛋白点被注释为家蚕微孢子虫的极管蛋白3(PTP3);另有6个假定蛋白,其中5个源自家蚕微孢子虫,1个源自桑尺蠖来源微孢子虫;其它蛋白点则分别被注释为ATP酶、铁氧还蛋白、ABC转运蛋白、解旋酶等,其中一个被注释为HrpA样解旋酶的蛋白点源自桑尺蠖来源微孢子虫,其余则源自家蚕微孢子虫。初步推测桑尺蠖来源微孢子虫由于极管蛋白3相对表达量偏低或缺少PTP3的部分肽段或其结构不同,影响到极管的弹射伸长,因而对家蚕的侵染力下降。
Through comparing proteomic profiles of Nosema bombycis and microsporidia isolated from Hemerophila atrilineata,the present study discussed the reason of virulence difference of these two microsporidia to silkworm（Bombyx mori）.Microsporidia of the two sources were first purified by Percoll method.Then,total proteins of microsporidia were extracted with glass beads breaking method.After protein concentration was measured by Bradford assay,the protein samples were subjected to two dimensional gel electropgoresis（2-DE）.Analysis and comparison to total protein 2-DE images of Nosema bombycis and microsporidia isolated from Hemerophila atrilineata using Imagemaster software revealed that the latter lacked 11 protein spots and the former lacked 2 protein spots.Mass spectrometric identification to these 13 protein spots displayed that one differential protein of about 40 kD was annotated as polar tube protein 3（PTP3） of Nosema bombycis.Besides,there were 6 hypothetical proteins,among which 5 were from Nosema bombycis and 1 from microsporidia isolated from Hemerophila atrilineata.Other protein spots were annotated as ATPase,ferredoxin,ABC transporter,helicase and so on respectively,among which HrpA-like helicase was from microsporidia isolated from Hemerophila atrilineata and the rest were from Nosema bombycis.It is preliminarily inferred that the polar tube extrusion in microsporidia isolated from Hemerophila atrilineata is affected because of low expression level of polar tube protein 3,or lack of partial peptide of PTP3,or different structure of PTP3,leading to a decline in its infectivity to silkworm.
出处
《蚕业科学》
CAS
CSCD
北大核心
2013年第3期537-542,共6页
ACTA SERICOLOGICA SINICA
基金
现代农业产业技术体系专项(No.CARS-22)
关键词
微孢子虫
家蚕
桑尺蠖
蛋白质组
双向电泳
基质辅助激光解析电离串联飞行时间质谱
极管蛋白
Microsporidia
Bombyx mori
Hemerophila atrilineata
Protein
Two dimensional gel electropgoresis
Matrix-assisted laser desorption-ionization of flight-time mass spectrometry
Polar tube protein
