《植物生理学报》 2011, 47(1): 63-68

丹参非特异性脂质转移蛋白基因(SmLTP1)的克隆及其表达分析

刘梅, 生华, 化文平, 储君, 王喆之*

药用资源与天然药物化学教育部重点实验室, 西北濒危药材资源开发国家工程实验室, 陕西师范大学生命科学学院, 西安710062

通信作者：王喆之；E-mail: zzwang@snnu.edu.cn；Tel: 029-85301260

摘　要：

对丹参 EST 序列进行 Blast 分析, 获得一个新的非特异性脂质转移蛋白基因, 命名为 SmLT P1 (GenB ank 注册号为 EF187461)。该基因 cDNA 全长 593 bp, 包含一个长为 357 bp 的开放读码框, 编码 118 个氨基酸。生物信息学结构分析表 明, 该蛋白具有植物 nsLTP 的典型结构, 即 4 对二硫键, 4 个 α- 螺旋, 1 个可结合和容纳脂肪酸分子的类似口袋状的疏水结 构。实时荧光定量PCR 分析结果表明, SmLTP1 基因在丹参不同组织器官中差异表达, 其表达受病原菌和茉莉酸甲酯的诱 导, 显示SmLTP1 基因在植物防御反应中发挥作用。

关键词：丹参; 非特异性脂质转移蛋白; 基因克隆; 生物信息学分析; 实时荧光定量PCR

收稿：2010-09-08   修定：2010-11-17

资助：国家自然科学基金(31000134)和陕西省科技研究发展(攻关)计划(2006K16-G15)。

Cloning and Expression Analysis of A Non-Specific Lipid Transfer Protein Gene(SmLTP1) from Salvia miltiorrhiza Bunge

LIU Mei, SHENG Hua, HUA Wen-Ping, CHU Jun, WANG Zhe-Zhi*

Key Laboratory of Ministry of Education for Medicinal Resources and Natural Pharmaceutical Chemistry, National Engineering Laboratory for Resource Developing of Endangered Chinese Crude Drugs in Northwest of China, College of Life Sciences, Shaanxi Normal University, Xi’an 710062, China

Corresponding author: WANG Zhe-Zhi; E-mail: zzwang@snnu.edu.cn; Tel: 029-85301260

Abstract:

By analyzing EST sequences of Salvia miltiorrhiza, we found a new gene which showed high homology with plant non-specific lipid transfer proteins and named as SmLTP1 (GenBank accession number: EF187461). SmLTP1 consists of 593 nucleotides, including a single 357-bp opening reading frame and encoding a 118-amino-acid peptide. Bioinformatics analysis showed that SmLTP1 had the typical structure of the plant non-specific lipid transfer protein, including 4 pairs of disulfide, 4 α-helices and 1 tunnel-like hydrophobic cavity which could contain lipid molecules. Quantitative RT-PCR analysis revealed that the gene expressed in different organs, and could be induced by methyl jasmonate (MeJA) and pathogen. These results showed that the SmLTP1 might be pathogen-responsive gene in plant defenses and involve in a complex gene regulation network.

Key words: Salvia miltiorrhiza; non-specific lipid transfer proteins; gene clone; bioinformatics analysis; quantitative RT-PCR