草珊瑚叶片全长cDNA文库构建及EST序列分析  

Construction of full-length cDNA library of Sarcandra glabra and sequences analysis on EST

作　　者：谢德金[1] 沈少炎 姚旺[1] 荣俊冬[1] 何天友[2] 陈礼光[1] 郑郁善[1,2] 

机构地区：[1]福建农林大学林学院,福建福州350002 [2]福建农林大学园林学院,福建福州350002

出　　处：《中草药》2016年第23期4235-4241,共7页Chinese Traditional and Herbal Drugs

基　　金：福建省科技重大专项资助(2004YZ02-05);福建省科技创新平台资助(2008Y2001)

摘　　要：目的构建草珊瑚Sarcandra glabra叶片全长c DNA文库,进一步发掘与草珊瑚代谢及抗逆相关的基因,为草珊瑚功能基因的发现提供实验基础。方法以草珊瑚的嫩叶为实验材料,利用SMART法(即Clontech公司SMARTer试剂盒)构建草珊瑚全长c DNA文库。利用ABI3730 DNA序列分析测序获得大量的EST序列,利用生物信息分析方法对EST序列进行功能注释。结果构建了草珊瑚叶片的全长c DNA文库,经过鉴定草珊瑚c DNA文库的文库滴度为1.14×107 cfu/m L,平均插入片段为1 000 bp。利用该文库测序了221个单克隆,共获得EST序列177条,拼接出151个单一序列;利用NCBI数据库进行同源比对分析,共有119条(79%)与已知基因有显著的同源性,进行GO功能注释,显示其表达术语涉及了细胞生长,信号转导,蛋白质合成、转录,抗逆反应以及能量代谢等。结论构建了符合要求的草珊瑚叶片全长c DNA文库,并对相关EST序列进行了生物信息学分析,为草珊瑚基因组学研究提供一定的参考。Objective Sarcandra glabra was recognized as an important research material attributing to its high medicinal value and economic value.However,little information was known about its genomics and regulatory pathway participating in reproductive development.For the first step to understand the molecular basis and further explore genes which related to metabolism and resistance in S.glabra.Methods A SMART full-length complementary DNA library from the leaves tissue was constructed and characterized to providing the experimental basis for discovery of functional genes of S.glabra.The assembly expressed sequence tag （EST） data were completed by ABI3730 DNA program.A high quality full-length cDNA library was constructed successfully from S.glabra leaves.Results The titer of library was 1.14×107 pfu/mL and the average length of inserted fragments was 1 000 bp.A total of 221 clones were sequenced from the cDNA library and obtained 177 EST sequences.The EST sequences were assembled into 151 unigenes including 12 contigs and 119 singletons （79%）.EST exhibited significant similarity with known putative functional nucleotide sequences in the GenBank database.These genes were mostly involved in cell development,signal transduction,protein synthesis,transcription,stress tolerance response,energy metabolism based on molecular function of GO annotation.Conclusion This report constructs a full-length-cDNA library and analyzes the bioinformatics of the related EST sequences,and then offers a reference to genomic research of S.glabra.

关 键 词：草珊瑚 全长CDNA 表达序列标签(EST) 基因注释 GO功能注释 

分 类 号：R282.12[医药卫生—中药学]