家蚕石蚕突变(st)近等基因系与其回交亲本的表皮蛋白质组学差异分析  

Comparative Proteomic Analysis of Epidermal Tissue Between Bombyx mori Near-isogenic Line of st Mutant and Its Backcross Parent

作　　者：杜畅[1] 汪婉琳[1] 王学杨[1] 张彦[1] 徐家萍[1] 

机构地区：[1]安徽农业大学生命科学学院,合肥230036

出　　处：《蚕业科学》2013年第3期486-491,共6页ACTA SERICOLOGICA SINICA

基　　金：国家高技术研究发展计划"863"项目(No.2011AA100306);安徽省国际合作项目(No.11030603028)

摘　　要：以家蚕石蚕突变(st)近等基因系C8和其回交亲本C108(正常体型)的5龄幼虫表皮组织为材料,运用双向电泳(2-DE)技术分析家蚕st突变体与正常蚕体的表皮蛋白质组成差异。从2-DE图谱中选取表达差异较明显的蛋白点,用基质辅助激光解析飞行时间质谱(MALDI-TOF MS)成功鉴定出13种蛋白质,其中酶类相关蛋白质占50%以上。通过Real-time PCR检测这些酶类蛋白质基因在C8和C108的5龄幼虫表皮组织的转录水平,发现谷胱甘肽S-转移酶σ1(GSTs1)、磷酸甘油酸变位酶(Pglym)、磷酸丙糖异构酶(TPi)3个酶类蛋白质基因的转录水平差异显著,其中Pglym和TPi在C8表皮中的转录水平明显高于在C108表皮中的转录水平,而GSTs1在C108表皮中的转录水平相对较高。初步推测这些差异蛋白质可能与st突变体形成和导致蚕体硬实的性状有关。Epidermal tissues of the 5th instar of silkworm（Bombyx mori） near-isogenic line C8 of st mutant and its backcross parent C108（normal larval body） were used as the material to investigate the proteomic differences between st mutant and normal larval epidermal tissues by means of two dimensional electrophoresis.From the differentially expressed proteins,13 protein spots with obvious expressional difference were successfully identified by MALDI-TOF MS,among which enzyme-related proteins accounted for more than 50%.Real-time PCR assay on transcriptional levels of these enzyme-related protein genes in epidermis of the 5th instar larva of C8 and C108 revealed that there was significant difference in the gene transcription level of GSTs1,Pglym and TPi.Among them,Pglym and TPi had significantly higher transcription level in epidermis of C8 than that in epidermis of C108,and GSTs1 had relatively higher transcriptional level in C108.It is suggested that these differential proteins are involved in the formation of st mutant and stonification of its larval body.

关 键 词：家蚕 石蚕突变体 差异蛋白质 双向电泳 基质辅助激光解析飞行时间质谱 基因转录 荧光定量PCR 

分 类 号：S881.2[农业科学—特种经济动物饲养] Q51[农业科学—畜牧兽医]