木薯钙/CaM调控类受体激酶基因MeCRLK1的克隆及生物信息学分析

Cloning and Bioinformatic Analysis of Cassava Calcium/CaM Regulatory Receptor Kinase Gene MeCRLK1

木薯作为热带亚热带植物对低温非常敏感。为研究木薯对低温敏感的分子机制,我们从低温信号传导途径中的感知方面展开研究。植物类受体激酶作为信号传导途径中的信号接收器,在植物响应低温信号刺激过程中起重要作用。因此本研究利用RT-PCR从木薯栽培种SC8中克隆了一个钙/钙调素调控类受体激酶基因,命名为Me CRLK1(Gen Bank Accession No.KP256022)并进行了系统的生物信息学分析。研究结果表明该基因CDS区为1 296 bp,编码一个431 aa的蛋白。该蛋白的分子式为C_(2118)H_(3391)N_(605)O_(636)S_(18),分子量约为48.08 k D,理论等电点为8.11。该蛋白预测具有多个不同的磷酸化位点,3个Ca M结合位点,4个紊乱区和1个球蛋白区。蛋白结构分析表明该蛋白具有一个跨膜区,一个S_TKc结构域,N末端没有信号肽。预测此蛋白可能亚细胞定位于膜上。Me CRLK1和At CRLK1蛋白序列分析表明,它们的相似性为79.8%,序列一致性为77.6%。二者序列的差异主要位于C末端。该研究结果为进一步验证Me CRLK1基因的功能奠定了基础。

As a tropical and subtropical plant, cassava is quite sensitive to low temperature. To study the molecular mechanism of low temperature sensitivity of cassava, we began to research from the perception of low temperature signal transduction pathway. As a signal receiver in signal transduction pathway, plant receptor kinase plays an important role in plant response to low temperature signal stimulation. Thus in this study, we cloned a putative calcium/calmodulin-regulated receptor-like kinase gene by RT-PCR from cassava cultivar SC8, named it as MeCRLK1 （GenBank Accession No. KP256022） and then carried out systematic bioinformatic analysis. The results indicated that the gene CDS sequence was 1 296 bp in length, encoding a protein of 431 amino acids. The protein molecular formula was C2118H3391N605O636518, molecular weight was about 48.08 kD, and theoretical pI was 8. 11. The protein was predicted to have multiple different phosphorylation sites, 3 CaM binding sites, 4 disorder areas, and 1 globulin domain. The protein structure analysis indicated that the protein sequence had one transmembrane domain, one S_TKc domain, but there was no signal peptide in N terminal. We predicted that this protein might subcellularly localize on the membrane. MeCRLK1 and AtCRLK1 protein sequence analysis revealed that the similarity of the two was 79.8%, and the sequence identify was 77.6%. The sequence differences between MeCRLK1 and AtCRLK1 was mainly at the C terminal. The study laid a foundation for the further validation of the function of Me CRLK1 gene.