摘要
目的 克隆人参Panax ginseng中质体ATP/ADP转运蛋白基因(ATP/ADP transporter protein,AATP),为深入研究其在人参中的生物学功能奠定基础。方法 利用人参14个组织部位转录组测序的转录组数据库,通过与NCBI下载的其他植物中AATP的m RNA序列进行本地Blast比对,从中筛选出质体ATP/ADP转运蛋白基因。利用PCR技术克隆获得该基因的全长,通过生物信息学软件分析该基因编码蛋白的信息。利用人参14个组织部位的转录组数据库分析该基因在人参不同组织中的表达模式,并利用实时荧光定量PCR检测茉莉酸甲酯处理条件下该基因的表达模式。结果 获得人参AATP1基因全长c DNA,命名为Pg AATP1,该基因长度为1 866 bp,编码621个氨基酸,蛋白质相对分子质量为67 897.23,等电点为9.58。该蛋白与其他物种中的质体ATP/ADP转运蛋白比较类似,分析发现Pg AATP1在人参中所有组织中表达量均比较高,在果肉和叶片中表达量最高。实时荧光定量PCR检测发现Pg AATP1基因在茉莉酸甲酯处理条件下表达持续上调。结论 获得Pg AATP1基因全长cDNA序列,该基因在淀粉合成旺盛的组织中表达量较高,且响应茉莉酸甲酯处理。
Objective To clone the full-length cDNA of the ATP/ADP transporter protein(AATP)genes in Panax ginseng to provide resources and some knowledge necessary for the further gene function study.Methods The mRNA sequence of the AATP genes in other plants were downloaded on the website of NCBI and used to perform local Blast alignment in the transcriptome of Jilin ginseng from 14 tissues.The AATP gene in Panax ginseng was cloned by PCR,and analyzed using bioinformatics software and online resources.The expression pattern of PgAATP1 gene in 14 tissues of Panax ginseng was analyzed using the expression profile of transcriptome and its expression level under methyl jasmonate was deceted by quantitative real-time PCR.Results A full-length cDNA sequence was successfully cloned from Panax ginseng and named as PgAATP1,which was 1866 bp in length and encoded 621 amino acids.The relative molecular mass of PgAATP1 protein calculated was 67 897.23 Da,and the isoelecric point calculated was 9.58.It was found that the protein was similar to the plastid AATP in other species.The expression of this gene was high in all tissues but higherin fruit flesh and leaf blade,and the expression of PgAATP1 gene was up-regulated by methyl jasmonate.Conclusion We have obtained the full-length of PgAATP1 gene.This gene expressed higher in tissues of vigorous starch synthesis and responsing to methyl jasmonate.
作者
陈静
李纯
肖逸凡
孙春玉
王艳芳
王康宇
赵明珠
王义
张美萍
CHEN Jing;LI Chun;Xiao Yi-fan;SUN Chun-yu;WANG Yan-fang;WANG Kang-yu;ZHAOMing-zhu;WANG Yi;ZHANG Mei-ping(Research Center for Ginseng Genetic Resources,Jilin Agricultural University,Changchun130118,China;College of Life Science,Jilin Agricultural University,Changchun130118,China;College of Chinese Medicinal Materials,Jilin Agricultural University,Changchun130118,China)
出处
《中草药》
CAS
CSCD
北大核心
2019年第18期4411-4418,共8页
Chinese Traditional and Herbal Drugs
基金
国家“863”计划项目(2013AA102604)
吉林省发改委-吉林省农产业创新专项资金项目(2016C04)
吉林省科技厅自然科学基金项目(20180101027JC)
吉林省科技厅自然科学基金项目(20190201264JC)
关键词
人参
质体ATP/ADP转运蛋白
生物信息学
表达分析
茉莉酸甲酯
Panax ginseng C.A.Meyer
ATP/ADP transporter protein
bioinformatics
expression profile analysis
methyl jasmonate
