三七香叶基香叶基焦磷酸合酶基因的克隆及表达分析-中国中药杂志.doc

 PAGE \* MERGEFORMAT 11 三七香叶基香叶基焦磷酸合酶基因的克隆及表达分析 闵丹丹1，唐美琼2，李刚2，屈啸声2，缪剑华2, * (基金项目:?广西科学研究与技术开发项目?(桂科重1298001-1-4,?桂科1);?广西自然科学基金项目(2013GXNSFBA019087);?广西药用植物园青年基金项目?(桂药基20).*第一通讯作者?Tel:?(0)86-0771-5601139,?E-mail:?mjh1962@.**第二通讯作者?Tel:?(0)86-0771-5602461,?E-mail:?lggxu07@. 作者简介：闵丹丹，TelE-mail:md126.com. 广西中医药大学，南宁 530001；2广西壮族自治区药用植物园，广西药用资源保护与遗传改良重点实验室，南宁 530001） 摘要 基于三七转录组测序结果，选择萜类代谢骨架上香叶基香叶基焦磷酸合酶基因(GGPPS)，结合RACE技术，克隆得到全长1203bp的cDNA序列，最长开放阅读框为1 035 bp，命名为PnGGPPS，GeneBank登录号为：KM486564，PnGGPPS基因全长2 370 bp，包含1个内含子和2个外显子，该基因编码344个氨基酸，与HYPERLINK javascript:showjdsw(showjd_0,j_0)蓖麻（Ricinus communis）、丹参（Salvia miltiorrhiza）等植物GGPPS的同源性达到73%以上，具有富天冬氨酸区等多个结构域，属于类异戊二烯合成酶超家族。实时荧光定量PCR结果显示，PnGGPPS在1-3年生三七的根、茎、叶和3年生的花等不同器官组织中均有表达，但以3年生叶中具有显著水平的高表达量，暗示其既具有调节三七生长发育的基本功能，也与三七叶绿素和类胡萝卜素的合成、叶绿体的发育、阴生适应性，以及品质的形成有关。 关键词：三七；PnGGPPS基因；实时荧光定量PCR；阴生习性 Cloning and Expression analysis of the GGPPS Gene from Panax notoginseng MIN Dan-dan1, TANG Mei-qiong2, MIAO Jian-hua2,*, LI Gang2, **, Qu Xiao-sheng (1Guangxi University of Chinese Medicine，Nanning 530000，China；2Guangxi Botanical Garden of Medicinal Plant，Guangxi Key Laboratory of Medicinal Resources Protection and Genetic Improvement，Nanning 530023，China) Abstract：According to the transcriptome dataset of panax notoginseng, the key geranylgeranyl pyrophosphate synthase gene (GGPPS) in terpenoid backbone biosynthesis was selected to be cloned. Using specific primer pairs combining with RACE (rapid amplification of cDNA ends) technique, the full-length cDNA sequence with 1203bp, which containing a 1035bp open reading frame, was cloned and named as PnGGPPS. The corresponding full-length DNA sequence contained 2370bp, consisted of 1 intron and 2 exons. The deduced protein PnGGPPS contained 344 amino acids and shared more than 73% identity with GGPPS from Ricinus communis and Salvia miltiorrhiza. PnGGPPS also had specific Aspartic acid enrichment regions and other conserved domains, which belonged to t