无籽刺梨(Rosa kweichonensis var.sterilis)RksAGL基因克隆及表达分析

Cloning and Expression Analysis of RksAGL Gene in Seedless Chestnut Rose(Rosa kweichonensis var. sterilis)

以无籽刺梨(Rosa kweichonensis var.sterilis)花芽提取的RNA为模板,采用同源克隆结合RACE技术克隆得到无籽刺梨AGL基因的c DNA全长,命名为Rks AGL。序列分析表明,c DNA全长1 089 bp,开放阅读框长度为747 bp,可编码248个氨基酸。编码蛋白分子质量预测为28.56 k D,等电点为9.40,无信号肽序列。分析表明,Rks AGL基因属于MADS-box基因家族C类基因,其编码的蛋白与其它植物AGAMOUS类蛋白具有较高的同源性。实时定量PCR分析表明,RksAGL基因仅在雄蕊和心皮中表达。

With the floral bud RNA as template, the full length c DNA of AGL in seedless chestnut rose(Rosa kweichonensis var. sterilis) was cloned through homology-based cloning approach and rapid amplification of c DNA ends(RACE) technique. The full-length c DNA(1 089 bp) of AGL gene was obtained, named Rks AGL, with an open reading frame(ORF) of 747 bp and encoding 248 amino acids. The relative molecular mass of Rks AGL calculated was 28.56 k D, the isoelectric point was 9.40, and there was no signal peptide in Rks AGL. The results indicated that Rks AGL gene belongs to C-type gene and the deduced protein of Rks AGL was high identity with other AGAMOUS proteins from different plant species. Real-time quantitative PCR analysis showed that Rks AGL was expressed during stamen, carpel and was not expressed during sepal and petal.