两个烟草YUCCA6基因的克隆及分析

Cloning and Analysis of Two YUCCA6 from Tobacco

为了研究烟草生长素合成关键基因NtYUCCA6,本研究通过同源克隆从烟草品种云烟87中克隆了2个YUCCA6基因:NtYUCCA6-1和NtYUCCA6-2。分析了NtYUCCA6-1和NtYUCCA6-2基因编码的蛋白质基本理化性质、二级结构等。保守基元分析表明烟草NtYUCCA6-1和NtYUCCA6-2蛋白含有FAD结合位点和还原型辅酶Ⅱ结合位点,可能烟草的两个YUCCA6基因都具有生物学功能。进化分析表明烟草YUCCA6蛋白与番茄YUCCA6蛋白亲缘关系最近。另外,本研究对克隆基因在烟草根、茎、叶及腋芽中的表达模式进行了q PCR检测,结果显示:两个基因主要都在根中表达,但NtYUCCA6-2在所检测的组织中都有表达,而NtYUCCA6-1在茎中基本不表达。试验结果将有助于研究烟草YUCCA6基因在烟草生长发育中的具体功能。

In order to understand the function of YUCCA6 in tobacco, two YUCCA6 s, designated as NtYUCCA6-1 and NtYUCCA6-2, were cloned from cultured tobacco variety: Yunyan 87 in this study by BLAST against the NCBI database using the nucleotide sequence of At YUCCA6. The basic protein analyses including the physical and chemical features, secondary molecular structure were performed. The conservative motif analysis revealed that both proteins have FAD binding sites and NADPH binding sites, implying that both genes function in tobacco. The evolutionary analysis showed that YUCCA6 s from tobacco were most closely related to that from tomato. In addition, both genes are expressed most abundantly in root but displayed different tissue expression patterns in other tissues. NtYUCCA6-2 was expressed in all tissues tested whereas NtYUCCA6-1 expression couldn't be detected in the stem. Results from this study might contribute the further functional characterization of both genes in tobacco.