烟草优质品种叶片全长cDNA文库的构建和质量分析  

Construction of full-length cDNA library from leaf of quality tobacco varieties and its characterization

作　　者：曾建斌[1] 陈顺辉[2] 陈华[1] 贺小彦[1] 姜宝杰[1] 张冲[1] 蔡铁城[1] 庄伟建[1] 

机构地区：[1]福建省作物分子与细胞生物学重点实验室,福州市金山福建农林大学350002 [2]福建省烟草专卖局烟草农业科学研究所,福州市北环中路133号350003

出　　处：《中国烟草学报》2012年第1期85-91,共7页Acta Tabacaria Sinica

基　　金：国烟科(2006)371;闽烟科(2008)13;福建省作物分子与细胞生物学重点实验室项目(2008J1033)

摘　　要：为研究优质烟叶的分子基础,以翠碧一号不同生长时期的叶片为材料,CTAB法提取总RNA,利用SMART技术合成全长cDNA,之后按经修改的Clontech技术成功构建了烟草叶片全长cDNA文库。经鉴定,初级文库库容为3.85×106个克隆,重组率为100%,重组子插入片段集中在750-2000 bp之间。随机挑取25个克隆测序后,经生物信息学分析表明,烟草上已经报道的基因占10条,11条为全长基因序列;20条序列有功能注释。综合分析表明,所构建的文库质量较高,达到了基因的分离、筛选和克隆的建库目的。Leaves of variety Cuibi 1 from different periods were used as material to extract total RNA to study molecular mechanism of quality leaf tobacco, eDNA was synthesized by SMART technique and a fulllength eDNA library was constructed by modified method of clontech technology.3.85×10^6, the recombinant rate was 100% with insert Results indicated that the primary library storage capacity was fragment concentrated in 750-2000 bp. randomly selected and sequenced and subjected to preliminary bioinformatics analysis. Ten of reported in tobacco while eleven with full-length sequence and twenty sequences with function Twenty-five clones were the sequences had been annotation. Results also indicated that the library was of high quality, which lay the foundation for further cloning and screening target genes from the library.

关 键 词：烟草 叶片 SMART技术 CDNA文库 

分 类 号：S572[农业科学—烟草工业] Q7[农业科学—作物学]