摘要
目的克隆灰毡毛忍冬突变型品种AP1基因(Lm-XL-AP1),并对其进行生物信息学和时空表达分析。方法通过RACE技术克隆Lm-XL-AP1基因全长,运用生物信息学的方法进行基因同源性和相似性比较分析,预测其编码蛋白,并对其进行各种理化性质分析。运用荧光定量PCR(q RT-PCR)检测该基因在灰毡毛忍冬突变型植株不同花期和不同器官的相对表达量。结果获得AP1基因,开放阅读框(ORF)长729 bp,编码242个氨基酸,与甘菊中MADS-box基因家族AP1基因相似性达80%,且包含MADS和K-box的保守序列。AP1蛋白无跨膜区域,定位于细胞核中;在灰毡毛忍冬突变型植株第6花期相对表达量最高,同时茎、叶中也有表达。结论成功从灰毡毛忍冬突变型植株总RNA中克隆到可能参与控制花器官表达的AP1基因。
Objective To clone the AP1（Lm-XL-AP1） gene from a Special Variant Varieties of Lonicera macranthoides ＂Xianglei＂, and to analyze its bioinformatics and spatio-temporal expression. Methods Amplifing the full length of Lm-XL-AP1 gene by RACE technique, using bioinformatics method to analyze homology and similarity of the gene, predicting the coding protein and analyzing the various physical and chemical properties. The expression of the gene in different parts of Lonicera macranthoides Special Variant Varieties was detected by fluorescence quantitative PCR（qRT-PCR）. Results The AP1 gene, containing a 729 bp ORF that encoding 242 amino acids, was cloned. And the similarity of the gene compared with the AP1 gene from the MADS-box gene family of Chrysanthemum lavandulifolium up to 80%（Containing a conserved sequence of MADS and K-box）. Without transmembrane domain, AP1 was located in cell nucleus. It is expressed in various organs of Lonicera macranthoides Special Variant Varieties. Conclusion For the first time, the AP1 gene which may be involved in the control of the expression of floral organ was cloned from the total RNA of Lonicera macranthoides Special Variant Varieties.
作者
彭美晨
刘湘丹
徐玉琴
王珊
刘畅宇
陈勋
周日宝
PENG Mei-chen;LIU Xiang-dan;XU Yu-qin;WANG Shan;LIU Chang-yu;CHEN Xun;ZHOU Ri-bao(College of Pharmacy, Hunan University of Traditional Chinese Medicine, Changsha 410208, China;Hunan Traditional Chinese Medicine Piece Standardization and Function Technology Research Center, Changsha 410208, China;The First Hospital of Hunan University of Traditional Chinese Medicine, Changsha 410007, China;The Second Hospital of Hunan University of Traditional Chinese Medicine, Changsha 410000, China)
出处
《中草药》
CAS
CSCD
北大核心
2018年第7期1652-1660,共9页
Chinese Traditional and Herbal Drugs
基金
国家自然科学基金资助项目(81203007
81673546)
湖南省自然科学基金资助项目(2017JJ3237)
湖南省教育厅青年基金项目(16B193)
湖南省教育厅(14K071)
湖南省中药学重点学科基金项目(zy201505)
湖南省"中药学"重点学科建设项目资助(湘教通[2011]76号)
国家中医药管理局"药用植物学"重点学科资助(国中医药发[2009]30号)
湖南中医药大学研究生省级创新课题
关键词
灰毡毛忍冬
Lm-XL-AP1基因
生物信息学
荧光定量
克隆
Lonicera macranthoides Hand. -Mazz.
Lm-XL-AP 1 gene
bioinformatics
fluorescence quantification
cloning
作者简介
彭美晨（1992-），女，硕士研究生，研究方向为中药资源与质量。Tel：15084736469E-mail：810759831@qq．com;通信作者：周日宝，男，博士，教授，主要从事中药资源质量与开发研究。Tel：（0731）88458233
