Wang Zhou-Yong, Jiang Chao, Yuan Yuan, Chen Ping
Institute of Chinese Materia Medics, China Academy of Chinese Medicinal Sciences, Beijing 100700, China.
Zhongguo Zhong Yao Za Zhi. 2013 Jan;38(1):32-6.
To clone IspE and IspH unigene from Lonicera japonica and its substitutes, and analyze their gene sequence, protein properties and transcriptional level.
IspE and IspH unigene ware obtained from the transcriptome dataset of L. japonica. Full-length cDNA of IspE and IspH were cloned from buds of L. japonica, L. japonica var. chinensis, L. hypoglauca and L. dasystyla using RT-PCR technology, and named as LJIspE, LHIspE, LJCIspE, and LDIspE, LJIspH, LJCIspH, LHIspH and LDI-spH, respectively. And we also predicted the structure and function of IspE and IspH proteins.
IspE contained an open reading frame that consisted of 1 221 bp, encoding one polypeptide with 422 amino acids. A complete open reading frame of IspH gene consisted of 1 380 bp and encoded 459 amino acids. Both IspE and IspH ware non-secreted proteins and localized in the chloroplast. Transcripted level of IspE and IspH in bud of L. japonica, L. hypoglauca and L. dasystyla was not significantly difference, but their transcripted level in L. japonica var. chinensis was significantly higher than that in L. japonica.
The clone of IspE and IspH will help for further research on the synthesis of terpenes, aroma and color.
从忍冬及其替代品中克隆异戊烯基二磷酸异构酶E（IspE）和异戊烯基二磷酸异构酶H（IspH）单基因，并分析其基因序列、蛋白质特性和转录水平。
从忍冬转录组数据集中获得IspE和IspH单基因。利用逆转录聚合酶链反应（RT-PCR）技术从忍冬、红腺忍冬、淡红忍冬和华南忍冬的芽中克隆IspE和IspH的全长互补DNA（cDNA），分别命名为LJIspE、LHIspE、LJCIspE、LDIspE以及LJIspH、LJCIspH、LHIspH和LDI-spH。并且我们还预测了IspE和IspH蛋白的结构和功能。
IspE包含一个由1221个碱基对组成的开放阅读框，编码一个含422个氨基酸的多肽。IspH基因的一个完整开放阅读框由1380个碱基对组成，编码459个氨基酸。IspE和IspH均为非分泌蛋白，定位于叶绿体。IspE和IspH在忍冬、淡红忍冬和华南忍冬芽中的转录水平无显著差异，但在红腺忍冬中的转录水平显著高于忍冬。
IspE和IspH的克隆将有助于进一步研究萜类、香气和色素的合成。
