核盘菌子囊盘cDNA文库的构建、 EST序列分析与SsMADS基因的筛选

J4 ›› 2010, Vol. 07 ›› Issue (4): 694-698.

核盘菌子囊盘cDNA文库的构建、 EST序列分析与SsMADS基因的筛选

刘金亮, 郭艳梅, 郑明, 张世宏, 潘洪玉

吉林大学植物科学学院, 长春 130062

收稿日期:2009-10-21 出版日期:2010-07-26 发布日期:2011-06-14
通讯作者: 潘洪玉 E-mail:panhongyu@jlu.edu.cn

Construction of cDNA Library from Apothecium of Sclerotiniasclerotiorum Identification of SsMADS Gene by EST Sequencing

LIU Jinliang, GUO Yanmei, ZHENG Ming, ZHANG Shihong, PAN Hongyu

College of Plant Sciences, Jilin University, Changchun 130062, China

Received:2009-10-21 Online:2010-07-26 Published:2011-06-14
Contact: PAN Hongyu E-mail:panhongyu@jlu.edu.cn

摘要/Abstract

摘要：

以核盘菌子囊盘为材料, 应用SMART(Switch Mechanism at 5′end of RNA Transcript)技术, 构建了高质量的核盘菌cDNA文库. 涂平板测定和酶切反应鉴定表明, 原始文库的滴度为4.11×105, 重组率达97.3%. 通过对文库克隆的序列测定和初步生物信息学分析, 共获得1 800条核盘菌有效表达序列标签(Expressed Sequence Tag, EST). 同源比对分析表明, 844条独立基因代表了242个已知基因, 其余602个EST为未知序列. 这242个已知基因大致可以分为3类： 78个可归属于生物学过程, 97个可归属于分子功能, 67个可归属于细胞组分. 其中, 在97个分子功能基因中筛选到1个与MADSbox具有较高同源性的基因（SsMADS）, 推测可能与核盘菌的有性生殖过程有关.

关键词: 核盘菌；子囊盘； cDNA文库, EST序列； SsMADS基因

Abstract:

Using total RNA from the apothecium of Sclerotinia sclerotiorum, a fulllength cDNA library was constructed by SMART(Switch Mechan
ism at 5′end of RNA Transcript) technology. According to the evaluation, the constructed library had a high titer of 4.11×105 and 97.3% recombinant percentage. 1 800 ESTs from the library were sequenced and compared with the sequences in the GenBank database. By the homologous analysis of 844 unigenes, 602 clones appeared to be completely unknown and were likely to represent newly described ge
nes, whereas 242 clones were identified based on matches to sequences in the database. These identified clones were categorized into three categories: 78 in biological process, 97 in molecular function, 67 in cellular component. In 97 molecular functional clones, a SsMADS gene was obtained which shared high identities with MADSbox and related to sexual reproduction of Sclerotinia sclerotiorum.

Key words: Sclerotinia sclerotiorum； apothecium； cDNA library, expressed sequence tag (EST)； SsMADS gene

中图分类号:

Q939.95

刘金亮, 郭艳梅, 郑明, 张世宏, 潘洪玉. 核盘菌子囊盘cDNA文库的构建、 EST序列分析与SsMADS基因的筛选[J]. J4, 2010, 07(4): 694-698.

LIU Jin-Liang, GUO Yan-Mei, ZHENG Meng, ZHANG Shi-Hong, BO Hong-Yu. Construction of cDNA Library from Apothecium of Sclerotiniasclerotiorum Identification of SsMADS Gene by EST Sequencing[J]. J4, 2010, 07(4): 694-698.