三七病程相关蛋白PR10-2基因的克

发布时间：2018-02-24 22:34

本文关键词： 三七根腐病病程相关蛋白原核表达　出处：《中国中药杂志》2017年16期 　论文类型：期刊论文

【摘要】：在转录组测序的基础上,采用逆转录聚合酶链式反应(RT-PCR)技术,从三七主根中分离到三七病程相关蛋白10(pathogensis related protein 10,PR10)基因,命名为PnPR10-2,测序结果表明该序列长465 bp,编码154个氨基酸。氨基酸序列同源性及系统发育树分析发现,PnPR10-2与人参的PR10-2蛋白同源性最高,具有典型的病程相关蛋白Bet v I保守结构域。构建了重组载体pET32a(+)-PnPR10-2,在宿主菌Escherichia coli BL21中诱导表达融合蛋白,优化诱导条件,PnPR10-2融合蛋白在E.coli BL21中以可溶性和包涵体蛋白2种形式大量表达。纯化上清中的重组蛋白,运用纸片法分析纯化重组蛋白的体外抑菌活性,其对三七根腐病病菌腐皮镰刀菌Fusarium solani和坏损柱孢菌Cylindrocarpon destructans具有一定的抑制作用,猜测该基因可能参与了三七抗根腐病的防御反应。
[Abstract]:On the basis of transcriptome sequencing, reverse transcriptase polymerase chain reaction (RT-PCR) technique was used to isolate the pathogenesis-related protein 10pathogensis related protein (PR10) gene from the main root of Panax notoginseng (Panax notoginseng). PnPR10-2 was named as PnPR10-2.The result of sequencing showed that the sequence was 465bpand encoded 154 amino acids. The homology of amino acid sequence and phylogenetic tree showed that PnPR10-2 had the highest homology with ginseng PR10-2 protein. The recombinant vector pET32a (PnPR10-2) was constructed to induce the expression of fusion protein in Escherichia coli BL21. PnPR10-2 fusion protein was expressed in E. coli BL21 in two forms: soluble protein and inclusion body protein. The recombinant protein in supernatant was purified. The antibacterial activity of purified recombinant protein in vitro was analyzed by disk method. It has a certain inhibitory effect on Fusarium solani and Cylindrocarpon destructans of Panax notoginseng root rot. It is speculated that this gene may be involved in the defense response of Panax notoginseng against root rot.
【作者单位】：昆明理工大学生命科学与技术学院;云南省三七资源可持续利用重点实验室;
【基金】：国家自然科学基金项目(31560351,31260297)
【分类号】：S435.672

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