Lv Haizhou, Zhang Xin, Liao Baosheng, Liu Wanjing, He Liu, Song Jingyuan, Sun Chao, Luo Hongmei, Chen Shilin
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China.
Institute of Chinese Materia Medica, China Academy of Chinese Medicinal Sciences, Beijing 100700, China.
Acta Pharm Sin B. 2015 Nov;5(6):583-9. doi: 10.1016/j.apsb.2015.09.002. Epub 2015 Oct 21.
We cloned and analyzed the two genes of the 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase (HDR) gene family from Huperzia serrate. The two transcripts coding HDR, named HsHDR1 and HsHDR2, were discovered in the transcriptome dataset of H. serrate and were cloned by reverse transcription-polymerase chain reaction (RT-PCR). The physicochemical properties, protein domains, protein secondary structure, and 3D structure of the putative HsHDR1 and HsHDR2 proteins were analyzed. The full-length cDNA of the HsHDR1 gene contained 1431 bp encoding a putative protein with 476 amino acids, whereas the HsHDR2 gene contained 1428 bp encoding a putative protein of 475 amino acids. These two proteins contained the conserved domain of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase (PF02401), but without the transmembrane region and signal peptide. The most abundant expression of HsHDR1 and HsHDR2 was detected in H. serrate roots, followed by the stems and leaves. Our results provide a foundation for exploring the function of HsHDR1 and HsHDR2 in terpenoid and sterol biosynthesis in Huperziaceae plants.
我们从蛇足石杉中克隆并分析了1-羟基-2-甲基-2-(E)-丁烯基-4-二磷酸还原酶(HDR)基因家族的两个基因。在蛇足石杉的转录组数据集中发现了两个编码HDR的转录本,分别命名为HsHDR1和HsHDR2,并通过逆转录-聚合酶链反应(RT-PCR)进行了克隆。对推测的HsHDR1和HsHDR2蛋白的理化性质、蛋白结构域、蛋白二级结构和三维结构进行了分析。HsHDR1基因的全长cDNA包含1431 bp,编码一个含有476个氨基酸的推测蛋白,而HsHDR2基因包含1428 bp,编码一个含有475个氨基酸的推测蛋白。这两种蛋白都含有1-羟基-2-甲基-2-(E)-丁烯基-4-二磷酸还原酶的保守结构域(PF02401),但没有跨膜区域和信号肽。在蛇足石杉的根中检测到HsHDR1和HsHDR2的表达最为丰富,其次是茎和叶。我们的研究结果为探索HsHDR1和HsHDR2在石杉科植物萜类和甾醇生物合成中的功能奠定了基础。