小麦条锈菌PsNCS1基因的克隆及转录表达特征

Cloning and transcriptional profiling of PsNCS1 from Puccinia striiformis f.sp. Tritici

摘要:

摘要：【目的】克隆小麦条锈菌神经钙离子感应蛋白基因PsNCS1，分析其在病菌不同发育时期的表达水平。【方法】利用文库筛选和RT-PCR技术克隆PsNCS1的cDNA序列，采用生物信息学技术预测分析该基因编码蛋白的保守结构域及基本特性，构建系统发育树；运用实时荧光定量RT-PCR技术分析PsNCS1在病菌不同生长发育时期的表达水平。【结果】PsNCS1全长cDNA为1007 bp（GenBank登录号GU134621），开放阅读框为573 bp，编码190个氨基酸，分子量为22.17 kDa, 等电点为4.

Abstract:

Abstract: [Objective] We cloned PsNCS1 encoding neuronal calcium sensor from Pst and analyzed its transcriptional profile. [Methods] A full-length cDNA of PsNCS1 was cloned by using RT-PCR in combination with cDNA library screening, the sequence was analyzed with different bioinformatic tools and the gene expression pattern was characterized via real-time RT-PCR. [Results] PsNCS1 (Genbank accession no. GU134621) encoded 190 amino acids, with a molecular weight of 22.17 KDa and a pI of 4.96. PsNCS1 contained four conserved EF-hand domains and was N-terminally myristoylated. Phylogenetic analysis indicated that PsNCS1 was highly similar to the NCS from Basidiomycetes and the highest similarity was with that from Puccinia graminis (96%). Real-time RT-PCR analysis indicated the amount of PsNCS1 transcripts of urediospores and of germinated urediospores were doubled or more comparing with those of fungal bodies at other different developmental stages. [Conclusion] PsNCS1 might be involved in the process of urediospore formation and germ tube elongation. The present results may provide basic data for further analysis of the role of PsCNS1 in pathogenesis process and calcium signaling.