橡胶草(Taraxacum kok-saghyz Rodin)小橡胶粒子蛋白基因的克隆与表达分析

Cloning and Expression Analysis of Tarexacunm kok-saglvg Rodin Rubber Small Rubber Particle Protein Gene

以橡胶草(Taraxacum kok-saghyz Rodin)为试材,在茉莉酸处理的根转录组数据分析的基础上,采用PCR的方法克隆获得小橡胶粒子蛋白(small rubber particle protein,SRPP)基因。通过生物信息学分析研究了该基因的特征,并检测了该基因的时空和原核表达情况。结果表明:SRPP3蛋白分子质量24.431 kDa,理论等电点(pI)4.64,属于酸性蛋白,二级结构以β-折叠为主,具有REF保守结构域,属于REF/SRPP家族成员之一。系统进化树分析显示TkSRPP3基因与短角蒲公英同源性最近;时空表达分析表明,TkSRPP3基因在六月龄的根中表达最高,推测该基因可能与天然橡胶的合成有关。TkSRPP3原核表达最佳诱导条件为浓度1 mmol·L^-1 IPTG诱导表达6 h。该研究为深入了解REF/SRPP家族中SRPP3在天然橡胶中的作用提供了参考依据。

Using Tarexacunm kok-saglvg Rodin as experimental material,based on jasmonic acid-treated root transcriptome data analysis,PCR was used to clone the Small Rubber Particle Protein(SRPP)gene,and the open reading frame was long 690 bp,encoding 229 amino acids.The bioinformatics,spatiotemporal expression analysis and heterologous expression analysis of the gene were studied.The results showed that the molecular weight of SRPP3 protein was 24.431 kDa,the theoretical isoelectric point(pI)was 4.64,which was acidic protein,and the secondary structure was mainlyβ-sheet.It had a REF conserved domain and was a member of the REF/SRPP family.Phylogenetic tree analysis showed that the TkSRPP3 gene was the most homologous to the short-horned dandelion;the spatio-temporal expression analysis showed that the TkSRPP3 gene was the highest in the roots of June age,and the prediction was related to the synthesis of natural rubber.The prokaryotic expression of TkSRPP3 was induced at a concentration of 1 mmol·L^-1 IPTG for 6 h,which was the best induction condition.The aim is to lay the foundation for a preliminary study of the role of SRPP3 in natural rubber in the REF/SRPP family.