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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。
The homologous cloning method and RACE technique were combined to study the proteome difference of Panax notoginseng and to identify the full-length c DNA of PR10-1 protein. The preliminary analysis of the function of this gene in Panax notoginseng. Sequencing results showed that the full-length cDNA of this gene was 863 bp in length and contained an open reading frame (ORF) of 465 bp encoding 155 amino acids. The sequence was named Pn PR10-1 and GenBank Accession No. KJ741402. Amino acid sequence homology and phylogenetic tree analysis showed that Pn PR10-1 had the highest homology with PR10-1 protein of ginseng and contained many conserved domains such as Bet-v-I family. Real-time quantitative PCR analysis showed that Pn PR10-1 had background expression in all tissues of Panax notoginseng for 1-3 years, suggesting that Pn PR10-1 is involved in biological processes such as growth and development and metabolic regulation. Pn PR10-1 is affected by root rot pathogen (Fusarium oxysporum) stress up-regulated expression, presumably Pn PR10-1 gene may be involved in anti-pandemic and other broad-spectrum disease-resistant defense response.