Cloning and analysis of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase genes HsHDR1 and HsHDR2 in Huperzia serrate

• Published in: Acta pharmaceutica Sinica. B Vol. 5; no. 6; pp. 583 - 589
• Main Authors: Lv, Haizhou, Zhang, Xin, Liao, Baosheng, Liu, Wanjing, He, Liu, Song, Jingyuan, Sun, Chao, Luo, Hongmei, Chen, Shilin
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier 01.11.2015
• Subjects: 1-Hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase; Bioinformatics analysis; Gene clone; Huperzia serrate; Original; Terpenoid; Bioinformatics analysis; CAS, cycloartenol synthase; Terpenoid; MVA, mevalonic acid; SQS, qualene synthase; FPS, farnesyl pyrophosphate synthase; 1-Hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase; Gene clone; ORF, open reading frame; HMBPP, 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate; IPP, isopentenyl diphosphate; SE, squalene epoxidase; HDR, 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase; MEP, 2-methyl-d-eryth-ritol-4-phosphate; Huperzia serrate; DMAPP, dimethylallyl diphosphate; DXR, 1-deoxy-d-xylulose-5-phosphate reductoisomerase
• ISSN: 2211-3835; 2211-3843
• DOI: 10.1016/j.apsb.2015.09.002

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 Hs HDR1 and Hs HDR2,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 Hs HDR1 and Hs HDR2 proteins were analyzed.The full-length c DNA of the Hs HDR1 gene contained 1431 bp encoding a putative protein with 476 amino acids,whereas the Hs HDR2 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 Hs HDR1 and Hs HDR2 was detected in H.serrate roots,followed by the stems and leaves.Our results provide a foundation for exploring the function of Hs HDR1 and Hs HDR2 in terpenoid and sterol biosynthesis in Huperziaceae plants.