马铃薯StPSKRs基因的克隆及其亚细胞定位分析
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国家自然科学基金(31601703);


Cloning and Subcellular Localization Analysis of Potato StPSKRs Gene
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    摘要:

    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StPSKR1和 StPSKR2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StPSKR1和StPSKR2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StPSKR1和StPSKR2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StPSKR1和StPSKR2基因。(2)StPSKR1位于马铃薯1号染色体上,cDNA全长3 042 bp,编码1 013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;StPSKR2位于7号染色体,cDNA全长3 135 bp,编码1 044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。

    Abstract:

    In this study, the fulllength cDNAs of two potato plant phytosulfokine receptor genes StPSKR1 and StPSKR2 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 StPSKR1 and StPSKR2 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 StPSKR1 and StPSKR2 genes. (2) StPSKR1 is located on chromosome 1 of potato. The fulllength cDNA is 3 042 bp, encoding 1 013 amino acids. The relative molecular mass of predicted protein is 112.16 kD, the theoretical isoelectric point is 6.27. StPSKR2 is located on chromosome 7, and the fulllength cDNA is 3 135 bp. 1 044 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.

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卢 瑶,胡金雪,金 鑫,等.马铃薯StPSKRs基因的克隆及其亚细胞定位分析[J].西北植物学报,2019,39(9):1521-1527

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  • 在线发布日期: 2019-10-24
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