香石竹减数分裂重组相关基因DcRAD51D克隆及表达分析

Cloning and expression analysis of DcRAD51D in meiosis recombination in Dianthus caryophyllus

RAD51基因在减数分裂过程中起着十分重要的作用,能够编码一种重组酶,在DNA单链入侵完整双链过程中起作用。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个减数分裂重组相关基因RAD51D的全长cDNA序列,命名为DcRAD51D(GenBank登录号MK733915)。该基因全长1375bp,包含1个编码327个氨基酸长为984bp的开放阅读框。蛋白序列比对显示,DcRAD51D基因编码蛋白包含walker motif A和B基序,与藜麦的遗传关系较近。荧光定量PCR结果显示,DcRAD51D的表达量随着减数分裂进程而呈递减趋势,在花粉母细胞时期表达量最高。研究推测,DcRAD51D基因可能在香石竹减数分裂过程中起重要作用。

RAD51 gene is very important for meiosis,and can encode a recombinase,which plays a role in the process of DNA single-strand invasion.In this study,a new DcRAD51 D gene(the accession number in GenBank was MK733915)was isolated from the flower of Dianthus caryophyllus by using RT-PCR and RACE approaches.The full length cDNA sequence of DcRAD51 D was 1375 bp,with an ORF of 984 bp encoded 327 amino acids.Protein sequence alignment showed that DcRAD51 D contained walker motif A and B,and had highly similarity with RAD51 D of Chenopodium quinoa.Real time qRT-PCR showed that the expression level of DcRAD51 D,with the development of meiosis,was obviously decreased,and got to the maximum at the microspore mother cell phases.It was speculated that the DcRAD51 D might play an important role in meiosis of Dianthus caryophyllus.