• Archives of Pharmacal Research
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• v.28 no.6
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• pp.722-729
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• 2005

With the aim to improve the specificity and to reduce the cytotoxicity of polyethylenimine (PEI), we have synthesized the conjugates of the branched PEI (25 kDa) with transferrin. The trans-ferrin-PEI (TP) conjugates with five compositions were synthesized using periodate oxidation method and confirmed by FT-IR spectroscopy and gel permeation chromatography. The free amine contents of TP conjugates, which were able to condense and deliver DNA, increased as the amount of PEI increased. TP/DNA polyplexes were characterized by measuring gel elec-trophoresis, ethidium bromide fluorescence quenching, particle size and zeta potential of complexes. Complete complexation of the polyplexes was observed above the N/P ratio of 5 in TP/DNA, and above 3 in PEI/DNA, respectively. The zeta potential of the complexes decreased as the amount of transferrin in TP conjugates increased. Transfection efficiency of TP conjugates was evaluated in HeLa cell and Jurkat cell systems. Among the five compositions of TP conjugates, TP-2 system mediated a higher $\beta$-galactosidase gene expression than PEI system in Jurkat cell which was known to express elevated numbers of transferrin receptors. From the results of the cell viability based on MTT assay, TP conjugates showed lower cytotoxicity com-pared with the PEI system. We expect that the TP conjugate can be used efficiently as a non-viral gene delivery vector.

• KSBB Journal
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• v.29 no.4
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• pp.244-249
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• 2014

Biosynthesis pathway of medium-chain-length (MCL) polyhydroxyalkanoates (PHA) from fatty acid ${\beta}$-oxidation pathway was constructed in recombinant Escherichia coli by introducing the Pseudomonas sp. 61-3 PHA synthase gene (phaC2) and the maoC genes from Pseudomonas putida, Sinorhizobium meliloti, and Ralstonia eutropha. The metabolic link between fatty acid ${\beta}$-oxidation pathway and PHA biosynthesis pathway was constructed by MaoC, which is homologous to P. aeruginosa (R)-specific enoyl-CoA hydratase (PhaJ1). When the E. coli W3110 strains expressing the phaC2 gene and one of the maoC genes from P. putida, Sinorhizobium meliloti, and Ralstonia eutropha were cultured in LB medium containing 2 g/L of sodium decanoate as a carbon source, MCL-PHA that mainly consists of 3-hydroxyhexanoate (3HHx), 3-hydroxyoctanoate (3HO) and 3-hydroxydecanoate (3HD), was produced. The monomer composition of PHA and PHA contents varied depending on MaoC employed for the production of PHA. The highest PHA content of 18.7 wt% was achieved in recombinant E. coli W3110 expressing the phaC2 gene and the P. putida maoC gene. These results suggest that MCL-PHA biosynthesis pathway can be constructed in recombinant E. coli strains from the b-oxidation pathway by employing MaoC able to supply (R)-3-hydroxyacyl-CoA, the substrate of PHA synthase.

• Korean Journal of Plant Resources
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• v.12 no.4
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• pp.253-259
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• 1999

In this study, three simple methods were established to confirm the transgenic potato plants. The leaf disc was used in the first method. After leaf discs of transgenic and non-transgenic potato were transfered into the liquid MS medium with bialaphos 5mg/l, 25 days, the chlorosis occurred in the non-transgenic leaf discs while it could not find in the transgenic leaf discs, In the second method, shoot tips of potato were transferred into MS medium supplemented with 0.5mg/l bialaphos and 0.6% agar. After 7-10 days, a lot of roots developed from the transgenic shoot tip, but the non-transgenic shoot tip was dead. The third method was using chlorophyll contents. Leaf discs were transferred into the liquid MS medium with bialaphos 0.5 mg/l. After 15 days, the content of chlorophyll A in transgenic plant was at least 2.5 times higher than in non-transgenic plant. In addition, the PAT enzyme activity were detected in the transgenic potato, but not detected in normal potato.

• Journal of Bio-Environment Control
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• v.24 no.3
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• pp.264-270
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• 2015

Tropical night phenomenon has been increasing due to global warming recently, it is expected that fruit quality of apples will decrease due to elevated night temperature condition. In the present study, fruit quality at maturity, periodic anthocyanin biosynthetic gene expression and sugar contents in leaves and fruit flesh were investigated to establish the physiological responses of 'Fuji' apples under high night temperature. The night temperature were treated with such as ambient (control), ambient $-4^{\circ}C$, and ambient $+4^{\circ}C$. After the treatment, high night temperature didn't affect fruit diameter, weight, and soluble sugar contents. Coloration of ambient $+4^{\circ}C$ was poor than that of control, however there was no significant difference between these genes expression of control and that of ambient $+4^{\circ}C$ treatment in the late coloration season. Increase of sorbitol and glucose contents at ambient $+4^{\circ}C$ in leaves were smaller than those at control, and then sorbitol and sucrose contents in fruit flesh at ambient $+4^{\circ}C$ were smaller than those at control. The cross section of leaves showed that there were no differences with the structure of parenchyma and epidermis tissues between the treatments, but starch granules in the palisade parenchyma cells decreased in high night temperature treatments. Consequently, high night temperature didn't affect the fruit quality, but changed sugar contents in leaves and fruit flesh, and suppressed coloration regardless of anthocyanin biosynthetic gene expression.

• Journal of Bio-Environment Control
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• v.25 no.4
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• pp.240-248
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• 2016

The objective of this study was to determine the effect of exogenous abscisic acid (ABA) on the red coloration and endogenous ABA contents of apple fruit skins. ABA and fluridone (an ABA synthetic inhibitor, FD) was sprayed on 'Hongro' apple fruit skins at 107 days after full bloom (DAFB). Visual coloration and hunter's color values were not affected by the ABA and FD treatments. Anthocyanin contents in fruit skins increased similarly to hunter $a^*$ values of fruit skins, but ABA and FD did not affect its accumulations. Liquid chromatography analysis revealed that endogenous ABA contents in control fruit increased at first and then decreased from 12 hours after the treatment. ABA treatment increased ABA contents in fruit skins from 2 hour after the treatment and it lasted until the end of the treatments. FD decreased ABA contents in fruit skins from 6 hours after the treatment. ABA treatment increased MdNCED2 (an ABA biosynthetic gene), MdACO1 (an ethylene biosynthetic gene), and MdCHS and MdDFR expressions. However, MdUFGT expressions were not affected by ABA treatment.

• Proceedings of the Korea Contents Association Conference
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• 2018.05a
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• pp.561-562
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• 2018

디지털 중합효소연쇄반응(Digital PCR)은 3세대 PCR로 명명하며, 1세대인 일반 PCR과 2세대인 정량 PCR(Real-time PCR)의 단점을 보완하여 개발된 방법이다. Digital PCR System은 소량의 PCR 반응을 10만개 이상의 반응통(wall)에 적재하는 방식의 나노유체칩에서 쪼개어 증폭시킨 후, target DNA를 계수한다. Target DNA의 증폭 여부에 따라 positive(1)와 negative(0)로 digital signal처럼 받아들여 계수하고, 포아송 분포를 통해 target DNA의 copy를 계산해 최종적으로 샘플 microlitr당 Copy수로 결과 값을 확인할 수 있다. 본 연구에서는 종(種)이 다른 동물의 조직이 서로 섞여있을 때 사람의 조직을 탐색하는 방법으로 유전자 증폭을 할 경우, digital PCR의 유효성에 대해 증명하였다.

• BMB Reports
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• v.29 no.4
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• pp.308-313
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• 1996

An 1.4 kb of the fad3 cDNA encoding microsomal linoleic acid desaturase catalyzing the conversion of linoleic acid (18:2, ${\omega}-6$) to linolenic acid (18:2, ${\omega}-3$) was introduced into tobacco plants by the Agrobacterium-mediated plant transformation, Among the transgenic tobacco plants conferring kanamycin resistance, five transformants showing increment in unsaturated fatty acid contents were selected and further analyzed for the transgenecity, In genomic Southern blot analyses, copy numbers of the integrated fad3 DNA in chromosomal DNA of the five transgenic tobacco plants were varied among the transgenic lines. By Northern blot analyses, the abundancy of the fad3 mRNA transcript directed by Cauliflower Mosaic Virus 35S promoter was consistent with the relative copy number of the fad3 DNA integrated in the chromosome of transgenic tobacco plants. When compared with the wild type, accumulation of linolenic acid in transgenic tobacco roots was elevated 3.7- to 4.7-fold showing a corresponding decrease in the linoleic acid contents; however, slight increments for linolenic acid were noticed in transgenic leaf tissues. These results indicated that the elevated level of fad3 expression is achieved in transgenic tobacco plants.

• Animal Bioscience
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• v.35 no.8
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• pp.1195-1204
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• 2022

Objective: The objective of this study was to investigate the in vitro fermentation profiles of different soybean oligosaccharides (SBOs) and their effects on skatole production and cecal microbiota of broilers. Methods: Five SBOs with varying main component contents were fermented using an in vitro batch incubation inoculated with broiler cecal microbiota. Gas production was recorded automatically, skatole, indole and short-chain fatty acids (SCFAs) were determined using high-performance liquid chromatography, and microbial changes were analyzed using 16S DNA gene sequencing. Results: The addition of SBOs increased (p<0.05) gas production, suggesting bacterial growth-stimulating activities. In addition, the concentrations of indole were significantly (p<0.05) decreased after SBO supplementation, and SBO III, with higher sucrose and stachyose contents, decreased (p<0.05) the skatole level. Our results also revealed that the fermentation of SBOs by cecal microbiota produced (p<0.05) SCFAs, which were dominated by propionic acid, butyrate acid and lactic acid compared to the control. In addition, SBO III increased (p<0.05) the abundance of Firmicutes and Subdoligranulum and decreased that of Bacteroides. Conclusion: These results suggest that SBOs with higher sucrose and stachyose contents are promising prebiotics in modulating gut microbiota and reducing odor emission in broilers.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.2
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• pp.165-169
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• 2005

This study was performed to offer effective basic data for selection and improvement of Korean dairy cattle through identifying distributional properties among candidate genes (bovine butyrophilin, signal transducers and activators of transcription 5a, and prolactin hormone). In this study, polymorphisms of candidate genes were identified and the relationships between loci and production traits of each gene were analyzed using frozen semen of Holstein bulls (19 proven and 77 candidates). In butyrophilin (BTN) locus, polymorphisms information contents (PIC) value of BTN2 (0.372) was higher than those of others (BTN1; 0.155, BTN3; 0.254, BTN4; 0.169). As a result of analysis of genotyping STAT5a, using single stranded conformational polymorphism (SSCP) method and microsatellite locus, PIC values were 0.189 and 0.457, respectively. And PIC value of prolactin hormone gene was 0.176. In the relationships between genotypes and production traits, BTN3 was associated with 305-day production traits (p<0.05). PTAs for B allele were such as 110.43, 88.28 and 75.25 in BTN1, 3, 4 and these values were higher than those of A allele, but in the case of BTN2, A allele with 154.19 was higher than that of B allele. The results obtained from using candidate genes may be used as an useful index for the genetic improvement of dairy cattle population in Korea, and further studies are needed.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.568-570
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• 2003

The Hyoscyamus niger hyoscyamine $6{\beta}-hydroxylase$ (H6H, EC 1.14.11.11) gene was introduced into the genome of a Scopolia parviflora by the binary vector system using the disarmed Agrobacterium rhizogenes strain KCTC 2703. Expression of H6H enzyme which are involved in alkaloids pathway by western blot analysis using proteins extracted from leaf, stem flower, branch root and main root were examined The enzyme expression was found only in the roots, with no expression in leaf, stem and flower. The alkaloids contents were the most higher in root and then leaf and stem has very small amount of alkaloid contents were analyzed by HPLC. The expression level of H6H in transgenic plants were two or more times than wild type plants. In transgenic plant which constitutively expresses H6H enzyme, high concentration of scopolamine was accumulated.