• Journal of Plant Biotechnology
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• v.35 no.4
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• pp.299-306
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• 2008

A modified method of Agrobacterium-mediated perilla transformation was developed using two selection markers of an antibiotics (either hpt or nptll) and an herbicidal (bar) gene. Perilla hypocotyl explants were cocultured with Agrobacterium tumefaciens EHA 105 strain harboring plasmid vector (either pMOG6-Bar or pCK-Bar) for three days, respectively. Primary shoots were selected with antibiotics of hygromycin (15 mg/L) or kanamycin (125 mg/L) and regenerated shoots were further selected with herbicide phosphinothricin (ppt,1.2 mg/L) to obtain authentic transformants. Roots were induced for the regenerated shoots on the MS medium without hormone and 80 putative transgenic plants were obtained. Transgene integration into perilla genome was confirmed by Southern blot and their expression was analyzed by Northern blot. T1 perilla seeds drived from To plants were tested 0.3% basta spray for identification of stable gene delivery to next generation.

• Journal of Applied Biological Chemistry
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• v.54 no.1
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• pp.26-32
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• 2011

To improve genetic transformation of Brassica napus winter cultivar 'Youngsan', factors influencing shoot regeneration and transformation from cotyledonary petioles were investigated. Shoot induction was enhanced in the combination of 0.5 mg/L NAA and 2~4 mg/L kinetin. Silver nitrate was essential for successful shoot regeneration, ranging from 5 to 9 mg/L. The addition of $GA_3$ promoted plant regeneration. Among the tested Agrobacterium strains, co-cultivation times, and antibiotic selection regimes, choice of appropriate Agrobacterium strain was the most critical factor for efficient transformation of B. napus cv. 'Youngsan'. The EHA105 succinamopine strain was the most efficient and the maximum transformation efficiency was 26.8%. Transgenic shoots were selected on 10 mg/L phosphinothricin (PPT) containing media. The transgenic plants expressing bar and gus genes were resistant for commercial herbicide "Basta" and stained with X-Gluc. Southern blot hybridization indicated that the presence of one to three gus gene copies per genome and inheritance of the gus gene into the $T_1$ generation.

• Journal of Plant Biotechnology
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• v.35 no.1
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• pp.69-74
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• 2008

This study aims to establish the efficient soybean transformation system and develop soybean [Glycine max (L.) Merill] transformants using cotyledonary node explants. The cotyledonary node of soybean were co-cultivated with Agrobacterium tumefaciens strains (KYRT1, EHA105). These strains contain the binary vector pCAMBIA3301 which carries a herbicide-resistant far gene. Korean cultivars (Danbaekkong, Eunhakong) and foreign cultivars (Jack, Peking) were the most efficient in regenerating cotyledonary node. Therefore, they were chosen for the transformation. Results showed that the T-DNA transfer reached up to 60% and transformation efficiency reached up to 3% in the cotyledonary node explants from Jack cultivar, co-cultivated with EHA105 strain. Histochemical GUS evaluation showed that 12 individual lines, transformed with the 현 gene, have positive response. The transformed soybeans have been confirmed in the $T_0$ generation through phenotypic assay using herbicide $Basta^{(R)}$ and Southern blot analysis.

• Journal of Plant Biotechnology
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• v.41 no.3
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• pp.159-167
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• 2014

In eukaryotes, proteins that are secreted into ER are post-translationally modified by N-glycosylation, the patterns of which are significantly different between plant and animal cells. Biotechnology industry has already produced a number of therapeutic glycoproteins in plant cells. However, the aberrant glycosylation of therapeutic recombinant proteins in plant systems can cause immune problems in humans. Therefore, it is important to develop strategies for producing non-glycosylated forms to preserve biological activity and native conformation by a peptide: N-glycanase (PNGase). In this study, we try to isolate PNGase T gene from tomato, which can use as a platform plant for biotechnology industry. We isolated a cDNA (GenBank Accession number KM401550) from tomato leaves with 1,767 bp, which encoded a polypeptide of 588 amino acids with a predicted molecular mass of 65.8 kDa. We also investigated the expression patterns of PNGase T during fruit ripening of tomato. The transcripts of PNGase T, which were constitutively induced in tomato fruit from green stage, were significantly increased and reached a peak at orange stage. After which, those transcripts were continuously reduced. The expression pattern of PNGase T was coincided well with transcripts profiles of metacaspase gene, LeMCA, and senescence-related gene members of ACC synthase, LeACS2, LeACS4, and LeACS6, for ethylene biosynthesis during fruit ripening. These results suggest that PNGase T is involved in a de-glycosylation process associated with senescence and fruit ripening.

• Journal of Plant Biotechnology
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• v.29 no.2
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• pp.85-92
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• 2002

This study has been focused on improving transformation efficiency of rice using Agrobacterium tumefaciens. We have demonstrated the effect of this system when the GPAT gene related to the cold-resistance was transferred by Agrobacterium tumefaciens in rice. Transformation conditions were modified using intron $\beta$-glucuronidase (GUS) expression as a reporter gene in the rice. In this study, mature seed-derived calli of rice (Oruza sativa L. cv. Dongjin) were pre-cultured for 3 days and then infected with Agrobacterium. When this infected calli were cultured in the dark for 10 days on co-cu]lure medium containing 50 mg/L of CaCl$_2$, 30 mg/L of acetosyringone, 2 mg/L of 2,4-D, 120 mg/L of betaine, high GUS expression was observed. In the present transformation system, the efficiency of transformation of GPAT gene was about 54%. Stable integration of GPAT gene into chromosomal DNA was proven by southern blot analysis of genomic DNA isolated from T$_{0}$ progenies. The progenies (T1 generation) derived from primary transformant of 5 lines were segregated with a 3 (resistant) : 1 (sensitive ratio) in medium containing hygromycin. This high frequency transformation system can be used as a useful tool in transformation of another monocotyledon.n.

• Journal of Life Science
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• v.23 no.4
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• pp.487-494
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• 2013

An AlaAT knock-out mutant (alaat) of rice (Oryza sativa L.) was isolated from T-DNA tagging lines and the genotypes of its progeny were determined with AlaAT1-specific primers. The alaat phenotypes showed decreased growth and grain yield when compared with control plants. The activity of AlaAT1 in the mutant plants was practically undetectable. The responses of alaat plants to growth under salt stress were compared with those of control plants by measuring chlorophyll fluorescence and the activities and mRNA expression of antioxidant enzymes. All abiotic stresses tested (salt, drought, and chilling) caused a similar decrease in chlorophyll fluorescence in both alaat and wild type plants. The activity of peroxidase (POX), an antioxidant enzyme, decreased following salt treatment of alaat plants, while control plant showed an increased activity. The mRNA levels for cAPX (cytosolic ascorbate peroxidase), POX2, and AlaAT were determined by RT-PCR following salt stress. No AlaAT1 mRNA was detected in alaat plants. The POX2 mRNA showed a slightly increased level in the wild type but was not detected in alaat plants, in agreement with the activity assays. The levels of cAPX mRNA were greatly increased in both the wild type and alaat plants. The salt stress effects on rice plant growth are therefore proposed to reflect a loss of function of AlaAT, which alters the activity and synthesis of antioxidant enzymes (especially peroxidases), rather than a direct effect on photosynthesis.

• Analytical Science and Technology
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• v.19 no.1
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• pp.107-114
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• 2006

Early diagnosis and medical intervention are critical for the treatment of patients with metabolic disorders. A rapid analytical method was developed for simultaneous quantification of organic acids and amino acids in urine without labor-intensive pre-extraction procedure showing high sensitivity and specificity. A new method consisted of simple two-step trimethylsilyl (TMS)-trifluoroacetyl (TFA) derivatization using GC/MS-selective ion monitoring (SIM). Filter paper urine specimens were dried under nitrogen after being fortified with internal standard (tropate) in a mixture of distilled water and methanol. Methyl orange was added to the residue as indicator reagent. Silyl derivative of carboxylic functional group was followed by trifluoroacetyl derivative for amino functional group. N-methyl-N-(trimethylsilyl-trifluoroacetamide) and N-methyl-bistrifluoroacetamide were consecutively added and heated for 15-20 min at $65^{\circ}C-70^{\circ}C$, for TMS-TFA derivative, respectively. This reactant was analyzed by GC/MS-SIM. Linear dynamic range showed 0.001-50 mg with the detection limit of (S/N=3) 10-200 ng, and the quantification limit of 80-900 ng in urine. Correlation coefficient of regression line was 0.994-0.998. When the method was applied to the patients 'urine, it clearly differentiated the normal from the patient with metabolic disorder. The study showed that the developed method could be the method of choices in rapid and sensitive screening for organic aciduria and amino acidopathy.

• Journal of Life Science
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• v.31 no.12
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• pp.1079-1087
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• 2021

To analyze gut microbiota of livestock in Korea and compare taxonomic differences, we conducted 16S rRNA metagenomic analysis through next-generation sequencing. Fecal samples from broiler chickens, pigs, and cattle were collected from domestic feedlots randomly. α-diversity results showed that significant differences in estimated species richness estimates (Chao1 and ACE, Abundance-based coverage estimators) and species richness index (OUTs, Operational taxonomic units) were identified among the three groups. However, NPShannon, Shannon, and Simpson indices revealed that abundance and evenness of the species were statistically significant only for poultry (broiler chickens) and mammals (pigs and cattle). Firmicutes was the most predominant phylum in the three groups of fecal samples. Linear discriminant (LDA) effect size (LEfSe) analysis was conducted to reveal the ranking order of abundant taxa in each of the fecal samples. A size-effect over 2.0 on the logarithmic LDA score was used as a discriminative functional biomarker. As shown by the fecal analysis at the genus level, broiler chickens were characterized by the presence of Weissella and Lactobacillus, as well as pigs were characterized by the presence of provetella and cattele were characterized by the presence of Acinetobacter. A permutational multivariate analysis of variance (PERMANOVA) showed that differences of microbial clusters among three groups were significant at the confidence level. (p=0.001). This study provides basic data that could be useful in future research on microorganisms associated with performance growth, as well as in studies on the livestock gut microbiome to increase productivity in the domestic livestock industry.

• The korean journal of orthodontics
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• v.36 no.4
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• pp.284-294
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• 2006

Objective: Activating mutations in the fibroblast growth factor receptor-2 (FGFR2) have been shown to cause syndromic craniosynostosis such as Apert and Crouzon syndromes. The purpose of this pilot study was to investigate the resultant phenotypes induced by the two distinctive bone-targeted gene constructs of FGFR2, Pro253Arg and Cys278Phe, corresponding to human Apert and Crouzon syndromes respectively. Methods: Wild type and a transgenic mouse model with normal FGFR2 were used as controls to examine the validity of the microinjection. Micro-CT and morphometric analysis on the skull revealed the following results. Results: Both Apert and Crouzon mutants of FGFR2 induced fusion of calvarial sutures and anteroposteriorly constricted facial dimension, with anterior crossbite present only in Apert mice. Apert mice differed from Crouzon mice and transgenic mice with normal FGFR2 in the anterior cranial base flexure and calvarial flexure angle which implies a possible difference in the pathogenesis of the two mutations. In contrast, the transgenic mice with normal FGFR2 displayed normal craniofacial phenotype. Conclusion: Apert and Crouzon mutations appear to lead to genotype-specific phenotypes, possibly causing the distinctive sites and sequence of synostosis in the calvaria and cranial base. The exact function of the altered FGFR2 at each suture needs further investigation.

• Journal of Mushroom
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• v.12 no.1
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• pp.41-51
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• 2014

Cauliflower mushroom widely known high concent of ${\beta}$-glucan for farm cultivation invigoration verified characteristics of mycelia growth, genetic diversity, resistance to Trichoderma by replacement culture with Trichoderma and growth characteristics of new variety crossbleeding strain. The result of replacement culture with Trichoderma for verification resistance about Trichoderma, 6951 (T. viride) strain did not show special change after formation of confrontation line and 6952 (T. spp.) strain was showed more formation of spore after formation of confrontation line. But 6426 (T. harzianum) strain found to encroach part of growth area of cauliflower mushroom mycelia. Among 10 kinds cauliflower mushroom strain, JF02-06 strain collected by Gurye, found did not spore of Trichoderma and thought to be resistant to Trichoderma. The result of crossbleeding after selected that mother strain good growth and formation of fruit body, verified good mycelia growth at JF02-47, 49 and 50 strain in Korean pine of wood-chip media. The result of gene sequence about ITS1, 5.8S and ITS4 for analysis of genetic diversity at crossbleeding strain, found high significance to other cauliflower mushroom in registered Genebank. The result of growth characteristic of spore and mycelia of cauliflower mushroom by observation microscope, size of spore showed water drop shape to major axis $6{\mu}m$ and minor axis $5{\mu}m$ and clamp showed 3 types in mycelia. The wide of mycelia was $3{\mu}m$. The characteristic of mycelia of cauliflower mushroom found to grow mycelia in clamp at approximately 50%. The growth speed of mycelia was $0.507{\mu}m/min$ and 2nd mycelia grown similar speed to mother mycelia at parallel with mother mycelia after growth speed at $0.082{\mu}m/min$. The formation of clamp made small clamp for 5 hours after shown transfer of electrolyte in mycelia inside. The septum formation started after 3 hours and then finally completed after 2 hours. In this study, strain of cauliflower mushroom verified resistance of Trichoderma, genetic diversity and characteristic of mycelia growth. Therefore, basic knowledge of cauliflower mushroom will improve and further contribute to development of mushroom industry.