• The Korean Journal of Physiology and Pharmacology
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• 제21권6호
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• pp.625-632
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• 2017

Familial Parkinson's disease (PD) has been linked to point mutations and duplication of the ${\alpha}$-synuclein (${\alpha}$-syn) gene. Mutant ${\alpha}$-syn expression increases the vulnerability of neurons to exogenous insults. In this study, we developed a new PD model in the transgenic mice expressing mutant hemizygous (hemi) or homozygous (homo) A53T ${\alpha}$-synuclein (${\alpha}$-syn Tg) and their wildtype (WT) littermates by treatment with sub-toxic (10 mg/kg, i.p., daily for 5 days) or toxic (30 mg/kg, i.p., daily for 5 days) dose of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Tyrosine hydroxylase and Bcl-2 levels were reduced in the ${\alpha}$-syn Tg but not WT mice by sub-toxic MPTP injection. In the adhesive removal test, time to remove paper was significantly increased only in the homo ${\alpha}$-syn Tg mice. In the challenging beam test, the hemi and homo ${\alpha}$-syn Tg mice spent significantly longer time to traverse as compared to that of WT group. In order to find out responsible proteins related with vulnerability of mutant ${\alpha}$-syn expressed neurons, DJ-1 and ubiquitin enzyme expressions were examined. In the SN, DJ-1 and ubiquitin conjugating enzyme, UBE2N, levels were significantly decreased in the ${\alpha}$-syn Tg mice. Moreover, A53T ${\alpha}$-syn overexpression decreased DJ-1 expression in SH-SY5Y cells. These findings suggest that the vulnerability to oxidative injury such as MPTP of A53T ${\alpha}$-syn mice can be explained by downregulation of DJ-1.

• Journal of Korean Medicine Rehabilitation
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• 제20권2호
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• pp.1-15
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• 2010

Objectives : This study was performed to evaluate the effects of root of Cibotii rhizoma(CR) ethanol extract on the tissue and neuronal damage of the spinal cord injury(SCI). Methods : SCI was induced by mechanical contusion following laminectomy of 10th thoracic vertebra in Sprague-Dawley rats. CR was orally given once a day for 7 days after SCI. Tissue damage and nerve fiber degeneration were examined with cresyl violet and luxol fast blue(LFS) histochemistry. HSP72(as neuronal damage marker), MAP2(as nerve fiber degeneration marker), c-Fos(immediate early gene), and Bax(pro-apoptotic molecule) expressions were examined using immuno-histochemistry. Individual immuno-positive cells expressing HSP72, MAP2, c-Fos and Bax were observed on the damaged level and the upper thoracic and lower lumbar spinal segments. Results : 1. CR reduced degeneration of nerve fibers and motor neuron shrinkage in the ventral horn of the lower lumbar spinal segment, but generally it did not seem to ameliorate the tissue injury following SCI. 2. CR reduced demyelination in the ventral and lateral funiculus of the lower lumbar spinal segment. 3. CR reduced HSP72 expression on the neurons in the peri-central canal gray matter adjacent to the damaged region. 4. CR strengthened MAP2 expression on the motor neurons in the ventral horn and on nerve fibers in the lateral funiculus of the lower lumbar spinal segment. 5. CR reduced c-Fos positive cells in the peri-lesion and the dorsal horn of the damaged level and in the ventral horn of the lower lumbar spinal segment. 6. CR reduced Bax positive cells in the peri-lesion and the dorsal horn of the damaged level and in the ventral horn of the lower lumbar spinal segment. Conclusions : These results suggest that CR plays an inhibitory role against secondary neuronal damage and nerve fiber degeneration. following SCI.

• Korean Journal of Plant Resources
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• 제21권5호
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• pp.388-394
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• 2008

This study was carried out to develop new cultivars using the $T_5$ generation of transformed rice by PCR analysis with DNA marker in each generation $(from\;T_3\;to\;T_5)$. In the previous study, we successfully developed the transgenic rice plants over-expressing the Arabidopsis $H^+/Ca^{2+}$ antiporter CAX 1 (accession no. U57411) gene. The calcium concentration in brown rice of transgenic plants was higher than that of donor plants, Iipum, and was selected 3 lines out of 25 lines at cultured GMO field. The major agronomic traits such as culm length, panicle length and panicle number of 3 lines at transgenic plants $(T_5)$ were similar to wild type. Also these lines appeared to have disease resistance to rice blast, cold resistance as compared with donor types. The grain shape was similar to donor plant, however, the 1000 grain weight of brown rice was different from transgenic plants. These finding would be used for basic data of new variety registration.

• Weed & Turfgrass Science
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• 제4권2호
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• pp.118-123
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• 2015

Metagenomics is a powerful tool to isolate novel biocatalyst and biomolecules directly from the environmental DNA libraries. Since the metagenomics approach bypasses cultivation of microorganisms, un-cultured microorganisms that are majority of exists can be the richest reservoir for natural products discovery. To discover novel herbicidal substances from soil metagenome, we established three easy, simple and effective high throughput screening methods such as cucumber cotyledon leaf disc assay, microalgae assay and seed germination assay. Employing the methods, we isolated two active single clones (9-G1 and 9-G12) expressing herbicidal activity which whitened leaf discs, inhibited growth of microalgae and inhibited root growth of germinated Arabidopsis seeds. Spraying butanol fraction of the isolated active clones' culture broth led to growth retardation or desiccation of Digitalia sanguinalis (L) Scop. in vivo. These results represent that the screening methods established in this study are useful to screen herbicidal substances from metagenome libraries. Further identifying molecular structure of the herbicidal active substances and analyzing gene clusters encoding synthesis systems for the active substances are in progress.

• Korean Journal of Poultry Science
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• 제36권4호
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• pp.301-309
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• 2009

The present study was conducted to determine the effect of energy restricted (ER) diet on the expressions of lipogenic genes in liver and myogenic genes in muscle tissue of broiler chickens. Energy restriction was accomplished by providing chicks with 70% (ER70) or 85% (ER85) energy level of control diet intake. Energy restricted groups of chickens were restricted for 7 days, starting at 8 days of age. Ad libitum feeding was resumed after the restriction period, and continued through the end of the experiment. The body weight of chickens in the restricted groups gained less during the energy restriction period (P<0.05). The body weight of the ER groups were similar to the control group during the re-alimentation period. However, the body weight of the ER70 group did not catch up with that of the control group by 35 days of age. The energy restrictions during early life of chicks decreased the contents of triglycerides and cholesterol in blood (P<0.05), but those were not different among treatments after re-alimentation. The genes of fatty acid synthase (FAS) and transcription factors including SREBP and PPARγ were down regulated by restriction regimen only in ER70 (P<0.05). However, those genes were not completely recovered after re-alimentation in ER70 group. The RNA expression levels of Myo-D, Myf-5 and myogenin in all treatment groups were decreased by restriction regimen when compared with control group (P<0.05). Myogenin was highly expressed after re-alimentation, but the other genes were not different among groups. These results suggest that ER85 group shows the best growth performance by re-alimentation and the higher muscle differentiation by expressing myogenin.

• The Korean Journal of Nuclear Medicine
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• 제38권4호
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• pp.294-299
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• 2004

Purpose: Both human NIS and mutant $D_2R$ transgenes are proposed as reporting system in transplanted cell tracking. Using hepatoma cell lines, we constructed a dual reporter system containing human sodium-iodide symporter (hNIS) and dopamine 2 receptor ($D_2R$) and compared its characteristics. Materials and Methods: The recombinant plasmid ($pIRES-hNIS/D_2R$) was constructed with IRES (internal ribosome entry site) under control of the CMV promoter $pIRES-hNIS/D_2R$ was transfected to human hepatoma SK-Hep1 cell line with lipofectamine. HEP-ND ($SK-Hep1-hNIS/D_2R$) cells stably expressing hNIS and $D_2R$ was established by selection with G418 for two weeks. RT-PCR was performed to investigate the expression of both hNIS and $D_2R$ genes. The expressions of hNIS and $D_2R$ were measured by $^{125}I$ uptake assays and receptor binding assays. Specific binding of $D_2R$ to $[^3H]spiperone$ was verified by Scatchard plot with (+) butaclamol as a specific inhibitor. $K_d\;and\;B_{max}$ values were estimated. The correlation between hNIS and $D_2R$ expression was compared by using each clone. Results: Similar quantities of hNIS and $D_2R$ genes were expressed on HEP-ND as RT-PCR assays. HEP-ND cells showed 30 to 40 fold higher radioiodine uptakes than those of parental SK-Hep1 cells. $^{125}I$ uptake in HEP-ND cells was completely inhibited by $KClO_4$, a NIS inhibitor Specific binding to HEP-ND cells was saturable and the $K_d\;and\;B_{max}$ values for HEP-ND cells were 2.92 nM, 745.25 fmol/mg protein and 2.91nM, 1323 fmole/mg protein in two clones, respectively. The radioiodine uptake by hNIS activity and $D_2R$ binding was highly correlated. Conclusion: We developed a dual positron and gamma imaging reporter system of hNIS and $D_2R$ in a stably transfected cell line. We expect that $D_2R$ and hNIS genes can complement mutually as a nuclear reporting system or that $D_2R$ can be used as reporter gene when hNIS gene were used as a treatment gene.

• Korean Journal of Plant Resources
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• 제15권3호
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• pp.177-187
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• 2002

In our experiment, selected mutants were used which showed not only the phenotype of a specific unpolished rice but also phenotypes of EM 40, LO 1050, and TAL 214. Reciprocal crosses between the mutants were conducted to select strains which would have more quantity of lipids than before. The constitution of fatty acid was also tested to figure out nutritional aspects of the mutants. In the crossing between EM 40 mutants and mutants (LO 1050) having a thick aleurone layer, the expression of EM 40 mutants has no relation with the thickness of the aleurone layer. And the lipid content of new F$_2$ strains through the crossing is 4.15 ％. The lipid content is larger than those of the parents including Kinmaze and in other crossings of this experiment. This is attributed to the fact that the new F$_2$ strains are the products of the crossing between genes responsible for the size of buds, where lipid is accumulated, and genes accountable for the thickness of the aleurone layer. In the crossing between EM 40 mutants and TAL 214 mutants, lipid content of the new F$_2$ strains is 3.8 ％, higher than 2.92 ％ of TAL 214 mutants. But the degree of lipid increase is smaller than in two other crossings. This is probably because genes expressing the phenotypes of TAL 214 affect the size of EM 40, which gets smaller. The aleurone layer of the new F$_2$ strains is 12 $\mu\textrm{m}$ thicker than the layer of TAL 214 mutants, but 6 $\mu\textrm{m}$ thinner than that of parents (LO 1050) having a thick aleurone layer. This seems to be affected by the size of a microscope. The phenotype of the new F$_2$ strains appears to be similar to that of TAL 214. The lipid content of the new F$_2$ strains is 3.85 ％, larger than 2.92 ％ of TAL 214 and 3.01 % of LO 1050. The increase may be due to the aleurone layer of LO 1050. And the size of the bud of the unpolished rice, though it is not big enough like that of LO 1050, seems to be affected by the accumulation of genes in the thick aleurone layer. The accumulation may contribute to the increase in the content of lipid. When it comes to the constitution of fatty acid, there is little difference between parents like Kinmaze and the new F$_2$ strains. But oleic acid increases while linoleic acid decreases. And the decrease in the linolenic acid seems to contribute to the increase in lipid content. This fact also raises the possibility that genes accountable for specific phenotypes could change the quality of rice if the genes are accumulated. Now, experiments on strains which have large lipid content in EM 40 type 1(ge-1, 3.68 ％), EM type 2(ge-2, 2.91 ％), thick aleurone layer(4.63 ％), and starch layer(3.44 ％) are under way to figure out the effects of gene accumulation. These experiments are likely to present the ways for increasing the lipid content.

• Journal of Plant Biotechnology
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• 제45권3호
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• pp.171-182
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• 2018

Papaya (Carica papaya L.) is one of the crops widely planted in tropical and subtropical areas. The papaya fruit has low calories and are plentiful in vitamins A and C and in minerals. A major problem in papaya production is a plant disease caused by the papaya ringspot virus (PRSV). The first PRSV-resistant GM papaya expressing a PRSV coat protein gene was developed by USA scientists in 1992. The first commercial GM papaya cultivars derived from the event was approved by the US government in 1997. Development of transgenic papayas has been focused on vaccine production and limited agricultural traits, including insect and pathogen resistance, long shelf life, and aluminum and herbicide tolerance. Approximately 17 countries, including the USA and China, produced transgenic papayas and/or commercialized them, which provoked studies on biosafety assessment and development of GM-detection technologies. For the biosafety assessment of potential effects on human health, effects of long-term feeding to model animals have been studied in terms of toxicity and allergenicity. Studies on environmental safety assessment include influence on soil-microbial biodiversity and transfer to soil bacteria of GM selection markers. Many countries, such as Korea, the European Union, and Japan, that have strict regulations for GM crops have serious concerns about unintended introduction of GM cultivars and food commodities using unauthorized GM crops. Transgene- and/or GM event-specific molecular markers and technologies for genomics-based detection of unauthorized GM papaya have been developed and have resulted in the robust detection of GM papayas.

• Journal of Plant Biotechnology
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• 제41권3호
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• pp.146-158
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• 2014

Camelina sativa that belongs to Brassicaceae family is an emerging oilseed crop. Camelina seeds contain approximately 40% storage oils per seed dry weight, which are useful for human and animal diets and industrial applications. Microsomal delta-12 fatty acid desaturase2 (FAD2) catalyzes the conversion of oleic acid to linoleic acid. The polymorphisms of FAD2 genes are correlated with the levels of oleic acids in seed oils. Microsomal delta-12 fatty acid desaturase2 (FAD2) catalyzes the conversion of oleic acid to linoleic acid. The polymorphisms of FAD2 genes are correlated with the levels of oleic acids in seed oils. In this study, three CsFAD2 genes (CsFAD2-1, CsFAD2-2 and CsFAD2-3.1) were isolated from developing seeds of Camelina sativa (L.) cv. CAME. The nucleotide and deduced amino acid sequences of three CsFAD2 genes were compared with those from dicotyledon and monocotyledon plants including Camelina cultivars Sunesone and SRS933. Three histidine motifs (HECGH, HRRHH, and HVAHH) required for FAD activity and a hydrophobic valine or isoleucine residue, which is a SNP (single nucleotide polymorphism) marker related with enzyme activity are well conserved in three CsFAD2s. The expressions of CsFAD2-1 and CsFAD2-3.1 were ubiquitously detected in various Camelina organs, whereas the CsFAD2-2 transcripts were predominantly detected in flowers and developing seeds. The contents of oleic acids decreased, whereas the amounts of linoleic acid increased in dry seeds of transgenic fad2-2 lines expressing each CsFAD2 gene compared with fad2-2 mutant, indicating that three CsFAD2 genes are functionally active. The isolated CsFAD2 genes might be applicable in metabolic engineering of storage oils with high oleic acids in oilseed crops.