家蚕细胞周期蛋白家族基因在5龄幼虫组织中的表达谱分析  

Expression profile analysis of the Cyclin gene in the silk gland cell of silkworm,Bombyx mori

作　　者：刘丽华[1] 沈卫德[2,3] 李兵[2,3] 王文兵[4] 

机构地区：[1]通化师范学院生物系,吉林通化134000 [2]苏州大学基础医学与生物科学学院,江苏苏州215123 [3]现代丝绸国家工程实验室,江苏苏州215123 [4]江苏大学生命科学研究院,江苏镇江212001

出　　处：《西北农林科技大学学报（自然科学版）》2012年第11期13-17,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家自然科学基金项目(31072086)

摘　　要：【目的】研究家蚕细胞周期蛋白家族基因(CyclinA、CyclinB、CyclinB3、CyclinE、CyclinL1)在丝腺及其他组织中的表达情况,为探讨细胞周期蛋白家族基因在细胞周期中的作用提供参考。【方法】取家蚕5龄3d幼虫的脑、脂肪体、中肠、丝腺、马氏管、精巢、卵巢及血液等组织,用Trizol提取各组织的总RNA并反转录合成cDNA。以Actin3为内参基因,合成的cDNA为模板,利用每个基因的特异性引物进行实时荧光定量PCR,检测细胞周期蛋白家族基因在转录水平上的表达情况。【结果】在丝腺中,CyclinE基因的表达量最高,CyclinA、CyclinL1基因表达较弱,而CyclinB、CyclinB3基因没有表达;在其他组织中,CyclinA、CyclinB、CyclinB3、CyclinL1基因均在精巢中高表达,而CyclinE基因在脂肪体中表达较高。【结论】CyclinE基因在家蚕丝腺细胞G1/S期起重要作用,CyclinB、CyclinB3基因在G2/M期起作用;CyclinA基因可能参与丝腺染色体的后期复制;CyclinA、CyclinB、CyclinB3、CyclinL1基因在家蚕精巢中的精母细胞发育形成精子进行分裂时起着重要作用。【Objective】 The research was to study the expression of cyclin family genes（CyclinA,CyclinB,CyclinB3,CyclinE,and CyclinL1） in gland and other tissues of silkworm,and to discuss the the role of cyclin family genes in the cell cycle.【Method】 The brain,fatbody,midgut,silk gland,malpighian tube,testis,ovarian and blood of 5 larvae at age of 3 day were collected.The total RNA was extracted with Trizol from tissues,then synthesized to cDNA.We used Actin3 as reference gene,the cDNA as template to detect the gene transcription level of cell cycle protein family in silkworm by real-time PCR with the specificity primer of each gene.【Result】 In silk gland,CyclinE gene expression was the highest,and CyclinA,CyclinL1 gene expression were relative weaker.There was no expression detected in CyclinB and CyclinB3 gene.For other tissues,there was a high expression of CyclinA,CyclinB,CyclinB3 and CyclinL1 in testis.CyclinE gene showed high expression in the fat body.【Conclusion】 CyclinE gene plays an important role in G1/S phase of silk gland cells,CyclinB,and CyclinB3 gene work in the G2/M period.CyclinA may participate in the later period of chromosome copy in silk gland.CyclinA,CyclinB,CyclinB3 and CyclinL1 play important roles in the division period when spermatocyte develops into sperm.

关 键 词：细胞周期蛋白 丝腺 表达谱 家蚕 

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]