月季花瓣丝氨酸蛋白酶基因RhSep1的克隆及生物信息学分析

利用RT-PCR技术,从切花月季品种"Samantha"中克隆丝氨酸蛋白酶基因RhSep1。利用生物信息学技术对所得到的丝氨酸蛋白酶RhSep1序列进行结构与功能预测。结果表明:RhSep1基因全长2 442 bp,开放阅读框编码769个氨基酸,推定该丝氨酸蛋白酶分子质量为80.48 kD。通过NCBI和MEROPS肽酶数据库等对Rh-Sep1进行Protein Blast,发现RhSep1具有肽酶S8₃家族SA的典型结构域Petidase_S8₃（序列为Tyr-108-Leu-341）和PA_subtilisin_like的结构域（序列为Tyr-348-Ile-474）。预测RhSep1可能具有信号肽、明显疏水区和典型跨膜区,同时RhSep1氨基酸序列中存在较多蛋白激酶C和酪蛋白激酶Ⅱ磷酸化位点、豆蔻酰化位点。这些活性位点往往与蛋白的磷酸化、G蛋白相互作用等信号转导事件有关,RhSep1可能存在复杂的蛋白水平调控机制。

Cloning and Bioinformatics Analysis of RhSep1 of Serine Protease from Rosa hybrida

The serine protease gene from Rosa hybrida "Samantha" was cloned by RT-PCR,and named as RhSep1.Bioinformatics approach was used to analyze and predict the function and domain of RhSep1.Results show that RhSep1 gene is 2 442 bp in total length,and the opening reading frame codes for 769 amino acids.The RhSep1’ molecular mass may be 80.48 kD.The RhSep1 was protein blast by NCBI and MEROPS database.Results show that there are two typical structural domains of Petidase_S8₃（the sequences is Tyr-108-Leu-341） and PA_subtilisin_like（the sequences is Tyr-348-Ile-474）.The predictive results of RhSep1 indicate that maybe there is a signal peptide,a sharp hydrophobic region and a typical transmembrane domain,and there are many function sites.The function sites are mainly Protein kinase C phosphorylation site,Casein kinase II phosphorylation site and N-myristoylation site.These function sites are always involved in protein phosphorylation,G-protein interaction and signal transduction,etc.According to bioinformatics analysis,the RhSep1 has a more complicated regulation mechanism in protein level.