菰黑粉菌Rak1同源基因UeRak1的克隆及表达分析

Cloning and expression analysis of Rak1 protein homology gene UeRak1 fromUstilago esculenta

采用PCR技术克隆获得一个编码7个WD-重复序列蛋白的基因UeRak1.UeRak1基因全长1 994bp,有1个974bp大小的内含子,开放阅读框1 020bp,编码339个氨基酸,UeRak1具有保守的WD40结构域,为Gβ同源蛋白,结构及聚类分析结果表明UeRak1与玉米瘤黑粉菌(Ustilago maydis)中的Rak1亲缘关系最近.实时荧光定量PCR结果显示UeRak1的表达量在细胞融合生长期急剧升高;当气生菌丝开始生长后表达量开始下降;在菰黑粉菌MT-1和T-1中UeRak1的表达趋势基本没有差别;但是MT-1菌株中UeRak1基因的表达量均不到T-1菌株的一半.表明UeRak1基因与菰黑粉菌细胞融合和菌丝生长具有紧密的联系.

The UeRakl gene, encoding a protein containing seven WD repeat domains, was cloned with PCR. The full length of UeRakl is 1 994 bp, with an open reading frame of 1 020 bp and an intron of 974 bp. It encoded 339 amino acids. This protein has a conserved WD40 domain and is a G~ homologous protein. Structure and cluster analysis results show that UeRakl has the closest relationship to RaM in Ustilago maydis. The real time fluorescence quantitative PCR results show that the expression of UeRakl increases rapidly during the cell fusion growth period but declines when the aerial mycelium begins to grow. Additionally, the expression patterns of UeRakl in MT 1 and T 1 type strains is similar. However, the UeRakl gene expression in the MT 1 type strain is less than half that of the T 1 type strain. All the results show that the UeRak 1 gene is closely related to the cell fusion and mycelial growth.