白木香AsNINJA1基因全长cDNA克隆及其在愈伤组织中的茉莉酸甲酯诱导表达  

Full-length cDNA cloning of AsNINJA1 gene in Aquilaria sinensis and MeJA-induced expression in callus

作　　者：廖永翠[1] 徐艳红[1] 张争[1,2] 隋春[1] 梁良[1,3] 魏建和[1,2] 

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所,北京100193 [2]中国医学科学院北京协和医学院药用植物研究所海南分所,海南省南药资源保护与开发重点实验室,海南万宁571533 [3]山东中医药大学药学院,山东济南250012

出　　处：《中草药》2014年第20期2968-2973,共6页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81173481,31100220,31000136);协和学者特聘教授[医科人发(2012)282号];科技部国家创新人才计划重点领域创新团队(2013);中组部“万人计划”(2014)

摘　　要：目的克隆国产沉香基原植物白木香Aquilaria sinensis互作蛋白(jasmonate Zim-domain,JAZ)基因(NINJA),为深入研究其在沉香倍半萜合成途径中的功能奠定基础。方法以白木香总RNA为模板,采用RACE法和RT-PCR方法,克隆白木香NINJA基因cDNA全长,并进行生物信息学的分析;采用实时荧光定量(qRT-PCR)方法分析经茉莉酸甲酯(MeJA)处理的白木香愈伤组织中AsNINJA1基因的表达模式。结果获得白木香NINJA基因全长cDNA,命名为AsNINJA1。序列分析表明,该基因的序列全长为1 982 bp,包含一个1 221 bp的开放阅读框,编码406个氨基酸,蛋白质相对分子质量为43 697,等电点为6.02。qRT-PCR结果显示,经MeJA处理4 h后AsNINJA1基因相对表达量最高,是对照(未经MeJA处理的白木香愈伤组织)的近100倍,随后显著下降。结论获得AsNINJA1基因全长cDNA序列,该基因对MeJA诱导极为敏感,且能在伤害早期响应。Objective To clone the full-length cDNA of interacting protein of JAZ（NINJA） gene in Aquilaria sinensis, and to provide the basic information for further study on gene function in sesquiterpenes biosynthesis pathway. Methods With the total RNA as template, the full-length cDNA of NINJA in A. sinensis was cloned through RACE technique and RT-PCR method. The bioinformatics of cloing NINJA gene was analyzed as well. The expression mode of this gene was with MeJA treatment in A. sinensis callus detected by qRT-PCR method. Results The full-length cDNA（1982 bp） of NINJA gene in A. sinensis, named as AsNINJA1 was obtained with an open reading frame of 1221 bp and encoding 406 amino acids. The relative molecular mass of AsNINJA1 protein calculated was 43697, and the isoelectric point was 6.02. The qRT-PCR results indicated that MeJA treatment could stimulate the increase of mRNA expression of AsNINJA1; There was a sharp rise at 4h with nearly 100 times higher than the control（without MeJA treatment）, then dropped significantly. Conclusion The full-length cDNA sequence of AsNINJA1 gene is obtained; AsNINJA1 is extremely sensitive to MeJA treatment, and responded to the early damage.

关 键 词：白木香 AsNINJA1基因 cDNA快速末端扩增 序列分析 表达分析 茉莉酸甲酯 

分 类 号：R282.12[医药卫生—中药学]