基于RNA-Seq技术检测分析家蚕卵巢发育高表达基因  

Determination and Analysis of Highly Expressed Genes During Ovary Development of Bombyx mori Based on RNA-Seq Technology

作　　者：宋作伟[1] 黄茉莉[1] 薛仁宇[1,2] 曹广力[1,2] 贡成良[1,2] 

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]苏州大学现代丝绸国家工程实验室,江苏苏州215123

出　　处：《蚕业科学》2013年第4期663-673,共11页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金项目(No.31072085);国家重大基础研究计划"973"项目(No.2012CB114600)

摘　　要：为了从分子水平解析家蚕卵巢发育和卵子发生的机制,对家蚕卵巢组织进行深度RNA测序,并对高表达基因进行KEGG通路富集分析。取5龄3 d、5龄6 d及化蛹2 d的家蚕卵巢分别抽提RNA,并合成cDNA,将3组样品混合后进行RNA-Seq测序,得到的总reads条带数为6 306 078,平均长度为100 bp,其中clean reads达到98.87%,测序饱和度良好,证明测序数据真实可靠。clean reads对应的基因数为9 609个。以家蚕Actin3基因为参照,计算所测基因的log2(Fold_change),将log2(Fold_change)>2的基因定义为高表达基因,共有66个基因为高表达基因。利用KEGG通路富集分析发现这些基因主要集中在能量代谢、蛋白质合成、信号转导等路径中。研究结果为理解家蚕卵巢发育及卵子的发生提供了新的线索。To understand the mechanism of ovary development and oogenesis of silkworm（Bombyx mori） at molecular level,deep RNA sequencing of silkworm ovary tissue was carried out and KEGG pathway enrichment analysis was conducted on the highly expressed genes.cDNA samples were respectively synthesized from total RNAs isolated from the ovaries of day 3 and day 6 larvae of the 5th instar and day 2 pupae.RNA-Seq sequencing was performed after mRNA samples of the above three origins were mixed.The obtained total reads were 6 306 078 with average length of 100 bp.The percentage of clean reads reached 98.87%,demonstrating that the saturation degree of sequencing was excellent and the data were realistic and reliable.The clean reads were mapped to 9 609 genes.The log2（Fold_change） values of the genes relative to Actin3 gene were calculated and the gene with log2（Fold-change）＆gt;2 was defined as highly expressed gene.As a result,66 genes were found to be highly expressed genes.KEGG pathway enrichment analysis displayed that these genes were mainly involved in energy metabolism,protein synthesis and signal transduction.These results provide new clues to understand ovary development and oogenesis of silkworm.

关 键 词：家蚕 卵巢 基因表达 RNA-Seq测序 KEGG分析 

分 类 号：S881.26[农业科学—特种经济动物饲养]