摘要
采用同源克隆法和RACE技术相结合,对三七根部蛋白质组差异进行研究,鉴定PR10-1蛋白相应的基因全长c DNA,初步分析该基因在三七中的功能。测序结果显示该基因序列全长863 bp,包含1个序列长465 bp编码155个氨基酸的开放阅读框,命名为Pn PR10-1,Gen Bank登录号KJ741402。氨基酸序列同源性及系统发育树分析发现,Pn PR10-1与人参的PR10-1蛋白同源性最高,含有Bet-v-I家族等多个保守结构域。实时荧光定量PCR分析显示,Pn PR10-1在1~3年生三七各组织中均有本底表达,暗示其参与生长发育、代谢调控等生物学过程;Pn PR10-1在根部受根腐病原菌(Fusarium oxysporum)胁迫上调表达,推测Pn PR10-1基因可能参与抗三七根腐病等广谱抗病防御反应。
With homology cloning approaches coupling with RACE(rapid-amplification of c DNA ends) techniques, the full-length coding sequence of pathogenesis-related protein PR10-1 with differential expression was cloned from the total RNA of the root of Panax notoginseng, and its function was explored furtherly. As a result, the longest 465 bp ORF(named as Pn PR10-1 with the Accession No. KJ741402 in Gen Bank) was detected from the cloned sequence with full-length of c DNA of 863 bp. The corresponding peptide encoded consisted of 155 amino acids, contained some domains such as Bet-v-I, and showed high similarity with that from Panax ginseng by analysis of phylogenetic trees created from the alignments. Real-time quantitative PCR showed that the expression of Pn PR10-1 gene was constitutive in different tissues of 1-3 year old plant, suggesting that it might be involved in growth, development, and secondary metabolism; yet it was up-regulated significantly with the infection of Fusarium oxysporum in root, suggesting that it might be involved in defense against many diseases including root rot in P. notoginseng.
出处
《药学学报》
CAS
CSCD
北大核心
2015年第2期227-232,共6页
Acta Pharmaceutica Sinica
基金
广西科学研究与技术开发项目(桂科重1298001-1-4)
广西药用植物园青年基金项目(桂药基20)
广西自然科学基金项目(2013GXNSFBA019087)
关键词
三七
病程相关蛋白
基因克隆
表达模式
防御反应
Panax notoginseng
pathogenesis-related protein
gene cloning
expression profile
defense response