核盘菌子囊盘cDNA文库的构建、EST序列分析与SsMADS基因的筛选  

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

作　　者：刘金亮[1] 郭艳梅[1] 郑明[1] 张世宏[1] 潘洪玉[1] 

机构地区：[1]吉林大学植物科学学院,长春130062

出　　处：《吉林大学学报（理学版）》2010年第4期694-698,共5页Journal of Jilin University:Science Edition

基　　金：国家自然科学基金(批准号:30971885);国家"十一五"科技支撑计划(批准号:2006BAD08A08);吉林省科技发展计划项目(批准号:20060544);吉林大学农学部重大项目启动基金;吉林大学基本科研业务费项目(批准号:200903376)

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4.11×105,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1800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.Using total RNA from the apothecium of Sclerotinia sclerotiorum,a full-length cDNA library was constructed by SMART（Switch Mechanism 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 genes,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.

关 键 词：核盘菌 子囊盘 CDNA文库 EST序列 SsMADS基因 

分 类 号：Q939.95[生物学—微生物学]