森林与环境学报2018，Vol.38Issue(1)：13-19,7.DOI:10.13324/j.cnki.jfcf.2018.01.003
PmACRE基因的克隆及遗传转化拟南芥
Isolation of PmACRE gene and its transformait on in Arabidopsis thaliana
摘要
Abstract
The Avr9/Cf-9 rapidly elicited ( ACRE) gene belongs to leucine rich repeats ( LRR) , which is the key gene involved in the process of protein-protein interaction and specific identification of pathogen elicitors and it plays a significant role in disease risistance.The Avr9/Cf-9 rapidly elicited gene from Pinus massoniana ( GenBank accession number: MF630966 ) was cloned through RT-PCR and named as PmACRE.The ACRE of P.massoniana, whose cDNA length is 862 bp, contained an open reading frame of 624 bp which encoded a polypeptide of 207 amino acids.The plant expression vector PFGFP Bar 137 was constructed, in which the PmACRE gene was driven by the constitutive promoter CaMV35S.The constructed expression vector was transformed into Agrobacterium tumefaciens AGL1 strain by the floral dip method.Leaf segments of Arabidopsis thaliana were inoculated with A.tumefaciens AGL1 containing plasmid PFGFP Bar 137-PmACRE.The transgenic lines were selected by ACRE and GFP gene PCR detection and the result showed that the target gene had been integrated into the genome of A.thaliana and the 4 positive plants have been obtained.The results of this study gave a foundation for the further analysis of the function of ACRE and revealing the mechanism on the P.massoniana in response to Bursaphelenchus xylophilus infestation.关键词
马尾松/PmACRE/基因克隆/载体构建/遗传转化Key words
Pinus massoniana Lamb./PmACRE/gene cloning/expression vector construction/genetic transformation分类
农业科技引用本文复制引用
李慧敏,谢婉凤,冯丽贞,陈慧洁,刘宏毅,叶小真..PmACRE基因的克隆及遗传转化拟南芥[J].森林与环境学报,2018,38(1):13-19,7.基金项目
福建省财政厅资助项目(K81139238K8113001A). （K81139238K8113001A）
