春性甘蓝型油菜BnMQ34基因的克隆及表达分析

Clone and Expression Analysis of BnMQ34 Gene in Spring Rapeseed

克隆春性特早熟甘蓝型油菜苗期表达的基因,并对其表达模式进行研究。用cDNA-AFLP技术筛选苗期特异性表达基因片段,根据差异片段设计特异性引物,用RACE技术扩增差异基因两端序列,并进行全长cDNA克隆。用RT-qPCR法进行表达分析。结果获得了一个苗期特异表达基因的cDNA全长序列455 bp,该基因编码一个由71个氨基酸残基组成的蛋白。该基因暂命名为BnMQ34。与油菜基因组序列比对结果显示,BnMQ34基因位于C4染色体上,由三个外显子和两个内含子组成。生物信息学分析显示,BnMQ34与甘蓝型油菜和白菜型油菜中报道的未诠释基因XM_013837282.1和XM_009127502.1的相似度达100%。油菜不同发育时期叶片中BnMQ34基因的表达量存在差异,苗期叶片中表达量最高。BnMQ34在春性特早熟甘蓝型油菜幼苗发育中可能起到重要的调节作用。

The purpose of this research was to clone the seedling specific expression gene and analyze its expression. The specific primers were designed according to a seedling specific expression gene fragment screened by cDNA-AFLP. Both end sequences of the gene were cloned using RACE technique. The full length cDNA was carried out and its expression pattern was studied by RT-qPCR. A full length cDNA in the gene of 450 bp was obtained, encoding a protein of 71 amino acid residues. This gene was tentatively designated as BnMQ34. Bioinformatics analysis showed that BnMQ34 gene was located on chromosome 4, consisting of three exons and one intron.It was showed that BnMQ3 had 100% similarity with unknown B. napus_013837282.1 and B. rapa XM_0091275-02.1. There were differences in the expression of genes in leaves at different developmental stages, and the highest expression was found in leaves at the seedling stage. It suggested that BnMQ3 gene might play a role in seedling development.