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拟南芥At PCa P1(protein bind calcium ions attaches to the plasma membrane,PCa P1)基因编码一个定位于细胞质膜上能结合钙离子且具有亲水性的跨膜蛋白,其参与协助了病毒粒子在植物胞间连丝中的运动,缺失该基因可导致芜菁花叶病毒在拟南芥中的累积量下降。本研究在甘蔗转录组数据库中搜索到与At PCa P1具有一定同源性的部分序列,并利用RACE(Rapid amplication of c DNA ends,RACE)技术从甘蔗品种台糖22的总RNA中克隆出完整的c DNA,命名为Sc PCa P1,Gen Bank收录号为KT891986。Sc PCa P1基因的c DNA全长为1 159 nt,5’端非编码区和3’端非编码区分别含有145 nt和312 nt,编码区包含一个702 nt的开放阅读框(Open reading frame,ORF),预测编码一个234个氨基酸的蛋白质,其分子量约为24.7 k D。同源性分析表明Sc PCa P1与拟南芥At PCa P1编码的蛋白的一致性为53.6%,与玉米和高粱的PCa P1蛋白一致性较高,分别是80.8%和85.2%。本研究进一步构建Sc PCa P1基因的RNAi(RNA interference)载体,用于后续研究Sc PCa P1在甘蔗抗病毒育种方面的作用及其他功能。
The Arabidopsis At PCa P1 (PCa P1) gene encodes a transmembrane protein that is localized to the plasma membrane of cells that binds to calcium ions and is hydrophilic, which is involved in assisting viral particles in plant Movement in the intercellular filaments, deletion of this gene, leads to a decrease in the accumulated amount of turnip mosaic virus in Arabidopsis thaliana. In this study, a partial sequence homologous to AtPaP1 was searched in the sugarcane transcriptome database and a complete cDNA was cloned from the total RNA of sugarcane variety TSS22 using RACE (Rapid Amplification of cDNA ends) c DNA, named Sc PCa P1, Gen Bank accession number KT891986. The full length cDNA of Sc PCa P1 gene was 1 159 nt. The 5 ’untranslated region and 3’ untranslated region contained 145 nt and 312 nt, respectively. The coding region contained a 702 nt open reading frame (ORF) ORF) predicted to encode a 234 amino acid protein with a molecular weight of about 24.7 kD. Homology analysis showed that the identity of Sc PCa P1 to Arabidopsis At PCa P1 protein was 53.6%, which was consistent with PCa P1 protein of maize and sorghum, which were 80.8% and 85.2% respectively. In this study, RNAi vector of Sc PCa P1 gene was further constructed to study the function and other functions of Sc PCa P1 in anti-virus breeding of sugarcane.