蓖麻RcDELLA基因的克隆及序列分析

Cloning and Sequence Analysis of RcDELLA Gene in Castor

DELLA家族蛋白是植物GA信号途径的重要阻遏蛋白,能抑制植物的生长和发育,使植株产生矮化的表型。为了明确DELLA基因在蓖麻中的作用和功能,对RcDELLA基因进行生物学信息分析。以蓖麻2129六叶期茎尖的cDNA为模板,根据NCBI公布的DELLA(XM_002533984.2)蛋白基因片段设计引物,克隆该基因,并对其进行生物信息学分析。克隆得到该基因的完整阅读框,该基因编码序列全长为1 701 bp,编码567个氨基酸,推测其分子量为62 550.40 ku,等电点为5.14。二级结构预测表明α-螺旋占40.4%,β-折叠占8.6%,无规则卷曲占51.0%。系统进化树结果表明,RcDELLA先与大戟科巴西橡胶树和麻疯树形成分支。通过PCR扩增得到了RcDELLA基因的完整阅读框,同源性比对结果显示,RcDELLA具有典型的DELLA结构域,与NCBI上公布的其他植物的DELLA蛋白序列具有高度的同源性。系统进化树结果显示,与麻疯树亲缘关系最近,符合进化关系。生物信息学分析将为进一步研究DELLA基因在蓖麻矮化过程中的作用和功能奠定理论基础。

DELLA family of proteins is an important repressor of plant GA signaling pathway, which can inhibit the growth and development of plants and make the plants have a dwarf phenotype. To clarify the function and function of DELLA gene in castor, conduct biological information analysis. Castor 2129 six-leaf stage shoot tip eDNA as a template, primers were designed according to the DELLA （XM_002533984.2） protein gene CDS published by NCBI, the gene was cloned and analyzed by bioinformatics. The complete reading frame of this gene was cloned,the full length of this gene was 1 701 bp, encoded 567 amino acids, molecular weight was 62 550.40 ku,isoelectric point was 5.14. The predicted secondary structure a-helix accounted for 40.4% ,13-fold accounted for 8.6% ,other non-random coil accounted 51.0%. Phylogenetic tree analysis showed that RcDELLA first formed a branch with Euphorbiaeeae rubber tree and Jatropha cur- cas. The complete reading frame of RcDELLA gene was amplified by PCR, the homology comparison showed that Rc- DELLA had the typical DELLA domain and had high homology with the DELLA protein sequence of other plants published on NCBI. Phylogenetie tree analysis showed that the genetic relationship with Jatropha curcas was theclosest and the result was consistent with species evolution. Bioinformatics analysis will lay the theoretical founda- tions for further study on the action and function of DELLA gene in castor dwarfing process.