家蚕细胞周期素E基因在5龄幼虫组织表达的剪切变体和表达谱特征  

Splicing Variants and Expression Profiles of CyclinE Gene in Tissues of the 5th Instar Bombyx mori Larvae

作　　者：刘丽华[1,2,3] 李兵[1,2] 沈卫德[1,2] 王文兵[4] 

机构地区：[1]苏州大学基础医学与生物科学学院,江苏苏州215123 [2]现代丝绸国家工程实验室,江苏苏州215123 [3]通化师范学院生物系,吉林通化134000 [4]江苏大学生命科学研究院,江苏镇江212013

出　　处：《蚕业科学》2011年第1期33-38,共6页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金项目(No.31072086);苏州市科技项目(No.SYN201009)

摘　　要：细胞周期素E（cyclinE）是调控细胞周期从G1期进入S期的关键因子。利用家蚕基因组数据库信息设计引物,克隆家蚕cyclinE在5龄幼虫不同组织表达的cDNA序列,获得该基因的6种不同剪切变体,这6种剪切变体前4个外显子和最后一个外显子都相同,变化主要集中在第5~7外显子处。同源性比对显示,不同物种来源的cyclinE周期蛋白盒区域具有较高的保守性,有35个氨基酸的共同保守序列,该区域可能也是家蚕cyclinE的特异识别并结合细胞周期素依赖蛋白激酶的功能区域。利用实时定量PCR分析显示,cyclinE在家蚕5龄第3天幼虫的中肠、脂肪体、丝腺、脑、马氏管、精巢、卵巢、血液等8种组织、器官中均有表达,并且在脂肪体中的表达相对较高,推测cyclinE基因在家蚕脂肪体细胞分裂过程中发挥重要作用。CyclinE is a key factor in the regulation of cell cycle from G1 phase to S phase. To study the expression of cyclinE gene in different tissues of silkworm （ Bombyx mori）, we designed primers by using information deposited in the silkworm genome database and cloned cyclinE cDNA sequences expressed in different tissues of the 5th instar silkworm larvae. As a result, 6 different splicing variants of this gene were obtained. The first 4 exons and the last exon of these 6 splicing variants were the same. Splicing variation mainly existed in the 5th to 7th exons. Homology comparison revealed that the cyclin box regions of cyclinE proteins from different species were highly conserved, among which 35 amino acids were identical. This region might also be the functional domain of silkworm cyclinE that involves in specific recognition and binding with cyclin-dependent protein kinase. Real-time quantitative PCR analyses showed that cyclinE was expressed in various tissues including midgut, fat body, silk gland, head, Malpighian tubule, testis, ovary and hemolymph of day 3 silkworm larvae of the 5th instar, and the expression level was relatively high in the fat body. It was suggested that silkworm cyclinE gene plays an important role in controlling the cell division in fat body.

关 键 词：家蚕 细胞周期素E基因 基因结构 剪切变体 组织表达 

分 类 号：S881.2[农业科学—特种经济动物饲养] Q78[农业科学—畜牧兽医]