家蚕翅原基表皮蛋白基因BmWCPs的生物信息学分析与激素诱导表达特征  

Bioinformatics Analysis and Ecdysone-induced Expression of Wing Cuticle Protein Genes BmWCPs in the Silkworm，Bombyx mori

作　　者：朱子丹[1] 章佳玲[1] 冯启理[1] 邓惠敏[1] 

机构地区：[1]华南师范大学生命科学学院、广州市昆虫发育调控与应用研究重点实验室,广州510631

出　　处：《蚕业科学》2016年第5期805-813,共9页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金重点项目(No.31330071),国家自然科学基金青年基金项目(No.31301918);国家重点基础研究发展计划“973”项目(No.2012CB114602)

摘　　要：翅原基表皮蛋白（WCPs）在昆虫翅的发育中起着非常重要的作用。为了研究家蚕翅原基表皮蛋白基因BmW CPs的表达是否与蜕皮激素（20E）的调节有关,用生物信息学方法分析BmW CPs基因的序列结构与系统进化特点,并通过半定量RT-PCR检测蜕皮激素对部分BmW CPs基因表达的诱导作用。11个BmW CPs基因成簇分布在家蚕基因组中,除BmW CP6和BmW CP10分别分布在第7号、第21号染色体外,其余的BmW CPs均分布在第22号染色体上;11个BmW CPs的核苷酸序列长度为588～2 670 bp,编码115～312个氨基酸,蛋白质分子质量12～35 kD,等电点4.30～7.45;11个BmW CPs基因的ORF内均有内含子插入,81.82%的基因含有2～3个内含子,内含子长度50～5 958 bp,在50～300 bp之间分布频率最高,占46%。将解剖获取的上蔟1 d蚕体的翅原基进行体外培养,并用浓度为2μmol/L的20E直接处理12 h后再常规培养18 h,提取翅原基总RNA并反转录合成cD NA,经半定量RT-PCR检测样品中BmW CPs的表达水平显著上调。依据上述研究结果初步认为,用蜕皮激素直接处理12 h后再常规培养18 h的脉冲作用,可诱导基因组中成簇分布的BmW CPs在翅原基组织中大量表达,以利于家蚕完成翅原基的变态发育。Wing cuticle proteins（ WCPs） play crucial roles in development of insect wings. To explore whether the expression of Bombyx mori wing cuticle protein genes（ BmW CPs） is associated with the regulation from ecdysone 20 E,bioinformatics methods were used to analyze the sequence structure and phylogenesis of BmW CPs and the ecdysone-induced expressional characteristics of BmW CPs were detected by semi quantitative RT-PCR. Bioinformatics analysis showed that the distribution of 11 BmW CPs formed as a cluster in the silkworm genome. Except that BmW CP6 and BmWCP10 were located on chromosome 7 and 21 respectively,the rest BmW CPs were all located in cluster on chromosome22. The nucleotide lengths of 11 BmW CPs were from 588 to 2 670 bp,encoding 115 to 312 amino acids. Their molecular weights were from 12 to 35 kD and their p Is were from 4. 30 to 7. 45. Gene structure analysis showed that all these genes had introns in their open reading frames. 81. 82% of BmW CPs contained 2 to 3 introns,with intron length ranging from 50 and 5 958 bp. Introns between 50 to 300 bp had the highest occurrence,accounting for 46% of total introns. After the wing discs ofsilkworm larvae at day 1 of cocooning stage were dissected and cultured in vitro,the wing discs were treated by ecdysone 20 E at the concentration of 2 μmol / L for 12 h and subject to routine culture for 18 h. Total RNA of wing discs was extracted and its c DNA was synthesized through reverse transcription. Compared to the control,the expression of BmWCPs increased significantly as shown by semi-quantitative RT-PCR detection. The above results initially demonstrate that high-level expressions of BmW CPs genes which exist as a cluster in the silkworm genome can be induced by ecdysone 20 E for 12 h followed by 18 h of routine culture,which greatly facilitate the completion of wing disc metamorphosis in silkworm.

关 键 词：家蚕 翅原基表皮蛋白基因 序列特征 诱导表达 蜕皮激素 

分 类 号：Q78[生物学—分子生物学]