title: Molecular cloning and characterization of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (CaHDR) from Camptotheca acuminata and its functional identification in Escherichia coli.
abstract: Camptothecin is an anti-cancer monoterpene indole alkaloid. The gene encoding 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (designated as CaHDR), the last catalytic enzyme of the MEP pathway for terpenoid biosynthesis, was isolated from camptothecin-producing Camptotheca acuminata. The full-length cDNA of CaHDR was 1686 bp encoding 459 amino acids. Comparison of the cDNA and genomic DNA of CaHDR revealed that there was no intron in genomic CaHDR. Southern blot analysis indicated that CaHDR belonged to a low-copy gene family. RT-PCR analysis revealed that CaHDR expressed constitutively in all tested plant organs with the highest expression level in flowers, and the expression of CaHDR could be induced by 100 microM methyl-jasmonate (MeJA), but not by 100 mg/L salicylic acid (SA) in the callus of C. acuminata. The complementation of CaHDR in Escherichia coli ispH mutant MG1655 demonstrated its function.
authors.affiliation: State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, Morgan-Tan International Center for Life Sciences, Fudan University, Shanghai, PR China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Wang
authors.forename: Yan
authors.initials: Y
authors.lastname: Pi
authors.forename: Rong
authors.initials: R
authors.lastname: Hou
authors.forename: Keji
authors.initials: K
authors.lastname: Jiang
authors.forename: Zhuoshi
authors.initials: Z
authors.lastname: Huang
authors.forename: Ming-Shiun
authors.initials: MS
authors.lastname: Hsieh
authors.forename: Xiaofen
authors.initials: X
authors.lastname: Sun
authors.forename: Kexuan
authors.initials: K
authors.lastname: Tang
mesh_terms: D000085:Acetates / Q000494:pharmacology; D015139:Blotting, Southern; D029769:Camptotheca / Q000201:enzymology*; D003001:Cloning, Molecular; D003517:Cyclopentanes / Q000494:pharmacology; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology; D035264:Flowers; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant / Q000235:genetics; D005816:Genetic Complementation Test; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D054883:Oxylipins / Q000494:pharmacology; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020156:Salicylic Acid / Q000494:pharmacology
chemical_list: D000085:Acetates; D003517:Cyclopentanes; D017931:DNA Primers; D018076:DNA, Complementary; D018744:DNA, Plant; D054883:Oxylipins; D012333:RNA, Messenger; C072239:methyl jasmonate; D010088:Oxidoreductases; D020156:Salicylic Acid