香石竹DcSKP1基因克隆及表达分析

Clone and expression analysis of DcSKP1 in Dianthus caryophyllus

SKP1基因是SCF E3泛素连接酶蛋白复合物的核心成分,参与多种生物过程。然而,香石竹SKP1基因还未被克隆,该文利用RT-PCR结合RACE技术,从香石竹(Dianthus caryophyllus)的花药中分离克隆了1个减数分裂相关基因SKP1的全长cDNA序列,命名为DcSKP1(GenBank登录号为MK931293)。结果表明:(1)DcSKP1基因cDNA全长序列为962 bp,含有1个长度为567 bp的ORF,该基因编码188个氨基酸。(2)蛋白序列比对显示,DcSKP1中存在一个高度保守TPEE基序,还具有Skp1_POZ结构域和Skp1结构域,并与拟南芥的SKP1聚集在一个分支上。(3)利用荧光定量PCR对香石竹DcSKP1基因表达模式进行研究,发现DcSKP1基因在各个组织部位都有表达,在花药中的表达量高于茎、叶组织,且在幼小的花药中表达量最高,随着花药发育的进程表达量下降。由此推测,DcSKP1基因可能在香石竹减数分裂中具有重要作用。

SKP1 gene is the core component of the SCF(Cul1-Rbx1-Skp1-F box)E3 ubiquitin ligase protein complex and is involved in many different biological processes.However,the SKP1 gene of carnation has not been cloned.In this study,a new DcSKP1 gene(the accession number in GenBank was MK931293)was isolated from the flower of Dianthus caryophyllus by using RT-PCR and RACE approaches.The results were as follows:(1)The full length cDNA sequence of DcSKP1 was 962 bp,and a ORF of 567 bp encoded 188 amino acids.(2)Protein sequence alignment showed that DcSKP1 had a highly conserved TPEE motif,Skp1_POZ domain and Skp1 domain,and clustered on a branch with Arabidopsis thaliana Skp1.(3)The DcSKP1 gene expression pattern of Dianthus caryophyllus was studied by qRT-PCR,and it was found that the DcSKP1 gene was expressed in all tissue sites,the expression level in anthers was higher than that in stem and leaf tissues,the expression level was the highest in young anthers,and the expression level decreased with the development of anthers.It is speculated that DcSKP1 gene may play an important role in carnation meiosis in D.caryophllus.