• Bulletin of the Korean Chemical Society
• /
• v.28 no.6
• /
• pp.965-969
• /
• 2007

The spectral properties, namely the circular dichroism, electric absorption and luminescence properties, of Λ- and Δ-[Ru(II)(1,10-phenanthroline)2benzodipyrido[b:3,2-h:2',3'-j]phenazine]2+ ([Ru(phen)2BDPPZ]2+) in the presence and absence of single stranded poly(dA) and poly(dT) were compared in this work. In the presence of single stranded DNAs, hypochromism in the absorption spectrum and significant changes in the circular dichroism spectrum in the ligand absorption band were apparent, indicating the strong interaction of the [Ru(phen)2BDPPZ]2+ complex with the single stranded DNAs. The luminescence intensity of the Ru(II) complex decreased stoichiometrically with increasing concentrations of the single stranded DNAs. All of these spectral changes were independent of the configuration of the Ru(II) complex and the nature of the DNA bases. Therefore, it is conceivable that both enantiomers of the [Ru(phen)2BDPPZ]2+ complex interact electrostatically with the negatively charged phosphate groups of DNA. However, the spectral properties of [Ru(II)(1,10-phenanthroline)3]2+ were not altered even in the presence of single stranded DNAs. Therefore, the size of the ligand involved in the interaction of the metal complex with the phosphate group of DNA may play an important role, even when the nature of the interaction is electrostatic.

• Asian-Australasian Journal of Animal Sciences
• /
• v.16 no.7
• /
• pp.1066-1070
• /
• 2003

In order to identify differentially expressed mRNAs (which represent possible candidates for significant phenotypic variances of muscle growth, meat quality between introduced European and Chinese indigenous pigs) in the longissimus dorsi muscle tissue between adult Duroc and Erhualian pigs, mRNA differential display was performed. Five 3' anchor primers in combination with 20 different 5' arbitrary primers (100 primer sets) were used and nearly 5,000 cDNA bands were examined, among which 10 differential display cDNAs were obtained, cloned and sequenced. Six of the 10 cDNAs showed similarity to identified genes from GenBank and the other 4 had no matches in GenBank. Differential expression was tested by Northern blot hybridization and could be confirmed for 2 cDNAs. The method used in this study provides a useful molecular tool to investigate genetic variation that occurs at the transcriptional level between different breeds.

• Journal of Life Science
• /
• v.8 no.3
• /
• pp.348-351
• /
• 1998

Common DNA fragments of the ${\beta}$-globin gene were observed from six races of the adult common carp: Hybrid-Yamato, Japanese wild type, Mirror, Suwa-Yamato, Scale German, and Saku-Yamato. Chromosomal DNAs isolated from the above six races were digested with restriction endonucleased EcoRI and PstI. The digested fragments were transferred onto nitrocellulose filter and hybridized with a probe of carp ${\beta}$-globin cDNA. Molecular sizes of the hybridized DNA fragments digested with EcoRI were 3.6Kb(Kilo base), 4.3Kb and 15Kb in Hybrid-Yamato, Japanese wild type, Mirror, Scale German and Saku-Yamato carp DNAs. In Scale German and Saku-Yamato carp DNAs, two and one more hybridized DNA fragments were observed, respectively. Molecular sizes of the hybridized DNA fragments digested with PstI were 2.2Kb, 6.5Kb, 7.8Kb and 9.2Kb in Hybrid-Yamato, 2.2Kb, 6.5Kb and 9.2Kb in Japanese wild type, 2.2Kb, 6.5Kb, 7.8Kb, and 13Kb in Mirror, 2,2Kb, 5,5Kb, 6.5Kb, 7.8Kb, 9.2Kb and13Kb in Scale German, and 2.2Kb, 5.5Kb, 6.5Kb, 9.2Kb and Saku-Yamato carp DNA. Therefore, depending on carps, three to six DNA fragments were hybridized with ${\beta}$-globin gene probe. Thus it indicated polymorphysm in the globin gene family of carp.

• Journal of Microbiology and Biotechnology
• /
• v.20 no.1
• /
• pp.187-192
• /
• 2010

In order to monitor the possibility of horizontal gene transfer between transgenic rice and microorganisms in a paddy rice field, the gene flow from a bifunctional fusion (TPSP) rice containing trehalose-6-phosphate synthase and phosphatase to microorganisms in soils was investigated. The soil samples collected from the paddy rice field during June 2004 to March 2006 were investigated by multiplex PCR, Southern hybridization, and amplified fragment length polymorphism (AFLP). The TPSP gene from soil genomic DNAs was not detected by PCR. Soil genomic DNAs did not show homologies on the Southern blotting data, indicating that gene transfer did not occur during the last two years in the paddy rice field. In addition, the AFLP band patterns produced by soil genomic DNAs from both transgenic and non-transgenic rice fields appeared similar to each other when analyzed by the NTSYSpc program. Thus, these data suggest that transgenic rice does not give a significant impact on the communities of soil microorganisms, although long-term observation may be needed.

• Journal of Plant Biotechnology
• /
• v.7 no.1
• /
• pp.51-55
• /
• 2005

Many cloning vectors in which cDNAs can be inserted to the sense orientation have been developed. Uni-ZAP XR vector (Stratagene) should contain clones that are oriented to sense direction with respect to T3 RNA polymerase primer. Unexpectedly, large portions of cDNAs in Chinese cabbage cDNA library showed unusual insertions, antisense orientation and a hybrid of two different clones. Using two clones, 4H03 and 53-B10, derived from different cDNA libraries, we proposed and demonstrated the possibility of unusual-construct formation by in vitro translation and northern blot analysis. The 4H03 clone was inserted with inverse direction, and its transcript and translation product could be produced by T7 RNA polymerase, indicating that this clone is definitely inserted into inverse orientation. The 53-B10 that contains two independent genes was turned out to be a hybrid in which two genes are inserted to opposite direction each other. All unusual constructs might be due to the presence of small fragments of DNA, like adapter. However, the mechanism underlined the formation of unusual constructs is still remain to be solved.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.11 no.5
• /
• pp.449-454
• /
• 2006

Here we present a sensitive DNA detection protocol using quantum dots (QDs) and magnetic beads (MBs) for large volume samples. In this study, QDs, conjugated with streptavidin, were used to produce fluorescent signals while magnetic beads (MBs) were used to isolate and concentrate the signals. The presence of target DNAs leads to the sandwich hybridization between the functionalized QDs, the target DNAs and the MBs. In fact, the QDs-MBs complex, which is bound using the target DNA, can be isolated and then concentrated. The binding of the QDs to the surface of the MBs was confirmed by confocal microscopy and Cd elemental analysis. It was found that the fluorescent intensity was proportional to concentration of the target DNA, while the presence of non-complementary DNA produced no significant fluorescent signal. In addition, the presence of low copies of target DNAs such as 0.5 pM in large volume samples up to 40mL was successfully detected by using a magnet-assisted concentration protocol which consequently results in the enhancement of the sensitivity more than 100-fold.

• Parasites, Hosts and Diseases
• /
• v.44 no.1 s.137
• /
• pp.15-20
• /
• 2006

Free-living amoebae of the genus Acanthamoeba are causative agents of granulomatous amebic encephalitis and amebic keratitis. Because the virulence of Acanthamoeba culbertsoni cultured in the laboratory is restored by consecutive brain passages, we examined the genes induced in mouse brain-passaged A. culbertsoni by differential display reverse transcriptase polymerase chain reaction (DDRT-PCR). Enhanced A. culbertsoni virulence was observed during the second mouse brain passage, i.e., infected mouse mortality increased from $5\%\;to\;70\%.$ Ten cDNAs induced during mouse brain passage were identified by DDRT-PCR and this was confirmed by northern blot analysis. BlastX searches of these cDNAs indicated the upregulations of genes encoding predictive NADH-dehydrogenase, proteasomal ATPase, and GDP-mannose pyrophosphorylase B, which have previously been reported to be associated with A. culbertsoni virulence factors.

• Bulletin of the Korean Chemical Society
• /
• v.35 no.4
• /
• pp.1105-1109
• /
• 2014

While single strand DNAs have been widely used for the scaffold of brightly fluorescent silver nanoclusters (Ag NCs), double strand DNAs have not been as successful. Herein, we report a novel synthetic approach for bright Ag NCs using branched double strand DNAs as the scaffolds for synthesis. X-shaped DNA (X-DNA) and Y-shaped DNA (Y-DNA) effectively stabilized Ag NCs, and both X-DNA and Y-DNA resulted in brightly fluorescent Ag NCs. The concentration and molar ratio of silver and DNA were found important for the fluorescence efficiency. The brightest Ag NC with the photoluminescence quantum efficiency of 19.8% was obtained for the reaction condition of 10 ${\mu}M$ X-DNA, 70 ${\mu}M$ silver, and the reaction time of 48 h. The fluorescence lifetime was about 2 ns for the Ag NCs and was also slightly dependent on the synthetic condition. Addition of Cu ions at the Ag NC preparations resulted in the quenching of Ag NC fluorescence, which was different to the brightening cases of single strand DNA stabilized Ag NCs.

• Journal of Microbiology and Biotechnology
• /
• v.29 no.6
• /
• pp.999-1007
• /
• 2019

Middle East respiratory syndrome coronavirus (MERS-CoV) was first identified in Saudi Arabia in 2012 and related infection cases have been reported in over 20 countries. Roughly 10,000 human cases have so far been reported in total with fatality rates at up to 40%. The majority of cases have occurred in Saudi Arabia with mostly sporadic outbreaks outside the country except for the one in South Korea in 2015. The Korean MERS-CoV strain was isolated from the second Korean patient and its genome was fully sequenced and deposited. To develop virus-specific protective and therapeutic agents against the Korean isolate and to investigate molecular determinants of virus-host interactions, it is of paramount importance to generate its full-length cDNA. Here we report that two full-length cDNAs from a Korean patient-isolated MERS-CoV strain were generated by a combination of conventional cloning techniques and efficient Gibson assembly reactions. The full-length cDNAs were validated by restriction analysis and their sequence was verified by Sanger method. The resulting cDNA was efficiently transcribed in vitro and the T7 promoter-driven expression was robust. The resulting reverse genetic system will add to the published list of MERS-CoV cDNAs and facilitate the development of Korean isolate-specific antiviral measures.

• Journal of fish pathology
• /
• v.34 no.1
• /
• pp.9-15
• /
• 2021

Rock bream (Oplegnathus fasciatus) is a highly valued aquaculture species in Korea. However, the aquaculture industry suffers huge economic losses due to rock bream iridovirus (RBIV) infection in summer. The objective of this study was to determine genetic diversity and relationships of DNAs isolated from two groups of rock bream after RBIV infection using five microsatellite (MS) markers. The first group of fish died early and the second group of fish died later after RBIV infection. In this experiment, 90 fish (5.1±1.0 cm and 4.1±1.3 g) were injected with 50 μl of RBIV (10⁴ TCID⁵⁰/ml) and maintained at 26℃ for 15 days. Genomic DNAs were extracted from fins of 20 fish that died earlier or later after RBIV infection. These DNAs were subjected to genotyping using five MS markers (CA-03, CA3-05, CA3-06, CA-10, and CA3-36). Of these markers, CA3-05 (early death group), CA3-06 (late death group), and CA3-36 (both early and late death groups) showed different alleles distribution rates. In-depth studies are needed to provide valuable information for selecting RBIV-resistant fish. In conclusion, microsatellite marker distribution pattern differences between early- and late- death groups of rock bream after RBIV infection showing different RBIV susceptibilities were determined using MS markers and genotyping. Results of this study suggest that MS markers could be used to facilitate the selection of RBIV resistant rock bream.