• Journal of Applied Biological Chemistry
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• v.48 no.2
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• pp.101-104
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• 2005

DXS catalyzes the first step of MEP pathway. Escherichia coli disruptants defective in dxs were constructed by insertional mutation and characterized. Selected disruptant, DXM3, was auxotrophic for DX or ME. Putative class 1 DXS ORF from Ginkgo biloba was shown to rescue DXM3 grown without DX or ME supplementation. The putative ORF was thus confirmed as DXS1. The disruptant was demonstrated to be useful for DSX screening.

• Korean Journal of Clinical Laboratory Science
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• v.50 no.2
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• pp.211-215
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• 2018

The aim of this study was to preserve the corticospinal tract during surgery and assess more accurately the motor performance in brain tumor patients around the motor cortex. TceMEP is not entirely reliable, even though there has been no change in waveforms due to a mixture of false positive and false negative signals. For a more detailed examination, DCS was employed to selectively stimulate the motor cortex. In both cases, the indications could find the region to which the cortex was responsible, and constantly check and examine the changes in amplitude, thereby preserving the motor pathway and performing surgery. On the other hand, patients who did not implement the DCS but did implement the TceMEP experienced a decrease in their postoperative motor performance. DCS is a very useful examination and it is a method that can reduce the post-surgery disorder that may occur in patients with the TceMEP in brain tumor surgery.

• BMB Reports
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• v.41 no.2
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• pp.112-118
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• 2008

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 ${\mu}M$ 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.

• Journal of Plant Biotechnology
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• v.39 no.3
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• pp.189-195
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• 2012

In plants, the fifth step of the plastidial 2-Cmethyl-D-erythritol 4-phosphate (MEP) pathway is catalyzed by 2-C-Methyl-D-erythritol 2,4-cyclodiphosphate synthase (MECP; EC: 4. 6. 1. 12), an enzyme proposed to play a key role in the regulation of isoprenoid biosynthesis. Here we report the isolation and functional characterization of a 823 bp Brassica rapa MECP (Brmecp) cDNA encoding a deduced polypeptide of 230 amino acid residues. Transcription levels of Brmecp were two-fold higher in petal compared to leaves. In addition, Brmecp expression in cabbage seedlings treated with ABA, $H_2O_2$ and drought was higher than control seedlings. These results were consistent with changes in chlorophyll contents in transgenic Arabidopsis. Thus, the Brmecp may contribute to the production of primary (chlorophylls and carotenoids) isoprenoid end-products in chloroplasts.

• Journal of Microbiology and Biotechnology
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• v.26 no.3
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• pp.441-451
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• 2016

Squalene is a linear triterpene formed via the MVA or MEP biosynthetic pathway and is widely distributed in bacteria, fungi, algae, plants, and animals. Metabolically, squalene is used not only as a precursor in the synthesis of complex secondary metabolites such as sterols, hormones, and vitamins, but also as a carbon source in aerobic and anaerobic fermentation in microorganisms. Owing to the increasing roles of squalene as an antioxidant, anticancer, and anti-inflammatory agent, the demand for this chemical is highly urgent. As a result, with the exception of traditional methods of the isolation of squalene from animals (shark liver oil) and plants, biotechnological methods using microorganisms as producers have afforded increased yield and productivity, but a reduction in progress. In this paper, we first review the biosynthetic routes of squalene and its typical derivatives, particularly the squalene synthase route. Second, typical biotechnological methods for the enhanced production of squalene using microbial cell factories are summarized and classified. Finally, the outline and discussion of the novel trend in the production of squalene with several updated events to 2015 are presented.

• BMB Reports
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• v.39 no.5
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• pp.502-510
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• 2006

2C-methyl-D-erythritol 2, 4-cyclodiphosphate synthase (MECPS, EC: 4.6.1.12) is the fifth enzyme of the non-mevalonate terpenoid pathway for isopentenyl diphosphate biosynthesis and is involved in the methylerythritol phosphate (MEP) pathway for ginkgolide biosynthesis. The full-length mecps cDNA sequence (designated as Gbmecps) was cloned and characterized for the first time from gymnosperm plant species, Ginkgo biloba, using RACE (rapid amplification of cDNA ends) technique. The full-length cDNA of Gbmecps was 874 bp containing a 720 bp open reading frame (ORF) encoding a peptide of 239 amino acids with a calculated molecular mass of 26.03 kDa and an isoelectric point of 8.83. Comparative and bioinformatic analyses revealed that GbMECPS showed extensive homology with MECPSs from other species and contained conserved residues owned by the MECPS protein family. Phylogenetic analysis indicated that GbMECPS was more ancient than other plant MECPSs. Tissue expression pattern analysis indicated that GbMECPS expressed the highest in roots, followed by in leaves, and the lowest in seeds. The color complementation assay indicated that GbMECPS could accelerate the accumulation of $\beta$-carotene. The cloning, characterization and functional analysis of GbMECPS will be helpful to understand more about the role of MECPS involved in the ginkgolides biosynthesis at the molecular level.

• Investigative Magnetic Resonance Imaging
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• v.4 no.1
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• pp.14-19
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• 2000

Purpose : This study was to present the functional brain mapping of both functional magnetic resonance imaging(MRI) and transcranial magnetic stimulation(TMS) in a case of schizencephaly. Materials and methods : A 28-year-old man, who had left hemiplegia and schizencephaly in right cerebral hemisphere, was exacted with both functional MRI and TMS. Motor function of left hand was decreased whereas right hand was within normal limit. For functional MRI, gradient-echo echo planar imaging($TR/TE/{\alpha}$=1.2 sec/90 msec/90) was employed. The paradigm of motor task consisted of repetitive self-paseo hand flexion-extension exercises with 1-2 Hz periods. An image set of 10 slices was repetitively acquired with 15 seconds alternating periods of task performance and rest and total 6 cycles (three ON periods and three OFF periods) were performed. In brain mapping, TMS was performed with the round magnetic stimulator (mean diameter; 90mm). The magnetic stimulation was done with 80% of maximal output. The latency and amplitude of motor evoked potential(MEP)s were obtained from both abductor pollicis brevis(APB) muscles. Results : Functional MRI revealed activation of the left primary motor cortex with flexion-extension exercises of healthy right hand. On the other hand, the left primary motor cortex, left supplementary motor cortex, and left promoter areas were activated with flexion-extension exercises of left hand. In TMS, magnetic evoked potentials were induced in no areas of right cerebral hemisphere, but in 5 areas of left corebral hemisphere from both abductor pollicis brevis. Latency, amplitude, and contour of response of the magnetic evoked potentials in both hands were similar. Conclusion : Functional MRI and TMS in a patient with schizencephaly were successfully used to localize cortical motor function. Ipsilateral motor pathway is thought to be secondary to reinforcement of the corticospinal tract of the ipsilateral motor cortex.

• Journal of Applied Biological Chemistry
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• v.58 no.3
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• pp.273-279
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• 2015

Phaeodactylum tricornutum is a model diatom that its genomic information and biological tools are well established. In this study, a gene encoding (E)-4-hydroxy-3-methylbut-2-enyl diphosphate reductase (PtHDR), a terminal enzyme of the methylerythritol phosphate pathway regulating chlorophyll and carotenoid biosynthesis, was isolated from P. tricornutum. The isolated gene was cloned into pPha-T1 vector containing fcpA promoter to prepare pPha-T1-HDR plasmid. As a positive control, pPha-T1-eGFP plasmid was constructed with egfp gene. Stable nuclear transformation was carried out with these plasmids by particle bombardment method and zeocin resistant colonies of P. tricornutum were selected on f/2 agar plate. In result, transformation efficiency was evaluated according to the amount of plasmid DNA coated with gold particles. Integration of introduced plasmids was confirmed with genomic DNA of each transformant by polymerase chain reaction. The eGFP fluorescence was visible in the cytoplasm, indicating that eGFP was successively expressed in P. tricornutum system. The transcript level of exogenous Pthdr gene was evaluated with the obtained transformants. The results presented here demonstrated that introduction of Pthdr gene into P. tricornutum chromosome succeeded and expression of PtHDR was enhanced under the fcpA promoter.

• KSBB Journal
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• v.21 no.2
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• pp.90-95
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• 2006

Escherichia coli harboring pAC-LYCO4 and pDdxs was used for lycopene production. Three wild type strains of E. coli OW1, MG1655, and W3110 were compared with DH5${\alpha}$ used before for lycopene production. Lycopene productivity of E. coli MG1655 was similar to DH5${\alpha}$ and the highest among those wild type strain. Therefore, MG1655 strain was used for random transposon and NTG mutagenesis to increase lycopene productivity. Through transposon mutation, five transposon mutants with increased lycopene productivity were obtained. It was found that genes knocked out by transposon insertion were treB in Tn1 mutant, B2436 in Tn2 mutant, and rfaH in Tn3, 4, and 5 mutants. Lycopene productivity was the highest in Tn4 mutant among the Tn mutants, which was 6-fold and 8-fold higher in lycopene concentration and content, respectively, in comparison with those obtained with wild type strain. NTG4 mutant was acquired with NTG mutation. The highest lycopene productivity of 6 mg/L and 4 mg/g DCW was obtained from the NTG4 mutant when arabinose of 0.013 mM was added for induction of dxs, rate-limiting gene of MEP pathway. The lycopene productivity of NTG4 mutant was increased 18-fold and 12-fold in lycopene concentration and content, respectively when comparing with the wild type strain.