基于转录组信息的艾纳香牻牛儿基牻牛儿基焦磷酸合成酶基因(BbGGPS)的克隆及序列分析

Analysis of geranylgeranyl pyrophosphate synthase(GGPS) sequence of Blumea balsamifera(L.)·DC based on transcriptome information

采用RT-PCR和RACE(Rapid amplification of c DNA ends)技术,从艾纳香(Blumea balsamifera L·DC)的叶片中克隆到二萜化合物合成的关键酶牻牛儿基牻牛儿基焦磷酸合成酶(Bb GGPS)基因。结果显示:Bb GGPS基因的c DNA全长1475 bp,包含开放阅读框(ORF)1002 bp,编码334个氨基酸;亚细胞结构定位于叶绿体,既非膜蛋白也非分泌性蛋白。疏水性分析显示,Bb GGPS是亲水性蛋白。同源性比对结果显示,Bb GGPS蛋白与其他植物中GGPS蛋白具有高度的相似性。系统发育分析表明,所有序列被聚为5大类,Bb GGPS与菊科植物刺菜蓟聚(Cynara cardunculus var)为一类,表明与其亲缘关系最近。

A geranylgeranyl diphosphate synthase gene,designated Bb GGPS,has been isolated from Blumea balsamifera（ L.） DC using reverse transcription polymerase chain reaction approach（ RT-PCR） and rapid amplification of c DNA ends（ RACE） methods. The results showed that the Bb GGPS c DNA had a full length of 1 475 bp,and contained an open reading frame predicting a polypeptide of 334 amino acids. Hydropathy and subcellular localization prediction showed that the Bb GGPS belongs to hydrophilic protein and is located in Chloroplasts. It is neither a membrane protein nor secretory protein. Bb GGPS protein showed a high similarity with other plant GGPS genes. Phylogenetic analysis indicated that all the amino acid sequence were divided into five categories and Bb GGPS was grouped with that of Cynara cardunculus var.