马铃薯StPSKRs基因的克隆及其亚细胞定位分析

Cloning and Subcellular Localization Analysis of Potato StPSKRs Gene

该研究采用同源克隆与PCR扩增方法,从马铃薯品种‘Desiree’中克隆植物磺肽素受体基因StPSKR 1和StPSKR 2的全长cDNA,并对其进行生物信息学分析及亚细胞定位分析,为深入研究StPSKR 1和StPSKR 2基因在马铃薯生长发育和生物胁迫中的作用提供理论依据。结果发现:(1)通过同源克隆与PCR扩增获得StPSKR 1和StPSKR 2的全长cDNA片段,并将其克隆到pGWB5-GFP载体;测序结果显示这2个基因编码的蛋白质与数据库给定的蛋白质序列保持一致,表明成功克隆到StPSKR 1和StPSKR 2基因。(2)StPSKR 1位于马铃薯1号染色体上,cDNA全长3042 bp,编码1013个氨基酸,预测蛋白相对分子质量为112.16 kD,理论等电点6.27;StPSKR 2位于7号染色体,cDNA全长3135 bp,编码1044个氨基酸,相对分子量为114.99 kD,理论等电点6.19。(3)生物信息学分析显示,StPSKR1和StPSKR2都属于跨膜蛋白。(4)亚细胞定位结果显示,StPSKR1和StPSKR2均定位于细胞膜上。

In this study,the full-length cDNAs of two potato plant phytosulfokine receptor genes StPSKR 1 and StPSKR 2 were cloned from potato“Desiree”,and their bioinformatic analysis and localization analysis were performed.These will contribute to study the roles of PSK signal in potato growth,development and response to biotic stress.The results showed that:(1)the full-length cDNA fragment of StPSKR 1 and StPSKR 2 genes were obtained by homologous cloning and PCR amplification,then cloned into pGWB5 vector.The protein sequence translated from the sequenced nucleotide results was consistent with that got from the database,indicating successful cloning of the StPSKR 1 and StPSKR 2 genes.(2)StPSKR 1 is located on chromosome 1 of potato.The full-length cDNA is 3042 bp,encoding 1013 amino acids.The relative molecular mass of predicted protein is 112.16 kD,the theoretical isoelectric point is 6.27.StPSKR 2 is located on chromosome 7,and the full-length cDNA is 3135 bp.1044 amino acids with a relative molecular weight of 114.99 kD and a theoretical isoelectric point of 6.19.(3)Bioinformatic analysis showed that both StPSKR1 and StPSKR2 belonged to transmembrane proteins,including extracellular LRR regions,transmembrane regions and intracellular kinase regions.(4)Subcellular localization analysis showed that StPSKR1 and StPSKR2 localized on the cell membrane.