丹参DHN1基因的克隆与序列分析

Cloning and sequence analysis of DHN1 gene from Salvia miltiorrhiza

丹参 SalviamiltiorrhizaBge. 是一种重要的药用植物.以组织培养2～13w的丹参幼苗为材料,构建丹参cDNA文库并进行大规模EST序列分析,所得序列经NCBI的BLAST工具分析,克隆号为rsmsxl-009377的序列与晚期胚胎丰富 LateEmbryogenesisAbundant 基因家族 中的成员有较高的同源性.根据其5′单向测序的结果设计引物 5′-GTGCGTAGACACATCGGTTC-3′ 继续向3′测序,得到一个全长969bp的序列,序列分析发现该序列包含一个长690bp的开放阅读框 ORF ,编码229个氨基酸,与NCBI注册的脱水素家族基因具有较高的同源性,且含有 LEA蛋白的特征序列,表明本基因可能是一种新的脱水素基因,命名为DHN1,并在GENEBANK上进行了注册,序列号为:AY695932.

Salvia miltiorrhiza Bge.is a sort of Traditional Chinese herbs.It's roots,called Dan-shen,have been formulated and used clinically for many years in China for the treatment of various diseases.This study regards 2～13 w tissue cultured Salvia miltiorrhiza Bge.seedlings as materials to construct the cDNA library and carry on extensive ESTs analysis.All the sequences are analyzed by BLAST tool of NCBI.The clone named rsmsxl-009377 showed high identity with late embryogenesis abundant typeⅡ proteins.Based on its 5′ sequences,we designed the primer '5′-GTGCGTAGACACATCGGTTC-3′' to continue the sequencing and have got a 969 bp sequence.Further analysis showed that this sequence has a 690 bp ORF encoding 229 amino acid-peptide and shows high identity with numerous dehydrin genes submitted to NCBI.Also the amino-acid sequence has all the characteristic sequences of LEA typeⅡ protein.We conjectured it's a new gene of LEA family and named it DHN1.It has submitted to GENEBANK and the sequence number is:AY695932.