• Journal of Periodontal and Implant Science
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• v.26 no.2
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• pp.365-375
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• 1996

5 serotypes(a, b, c, d, e) of Actinobacillus actinomycetemcomitans showed distinct hybridization patterns(DNA fingerprinting patterns) when the bacterial DNA were hybridized with randomly cloned 4.7-Kb sized DNA probe. The sizes of hybridized bands in each serotypes were different among serotypes and represented unique patterns of hybridization with the probe used. The serotype a showed two bands of fingerprinting patterns: 23.1 kb and 2.5 kb respectively. Serotype b and c showed single band: 6.6 kb and 9.5 kb, respectively. Serotype d and e showed two bands of hybridization: 23.1 kb and 2.8 kb, and 23.7 kb and 2.1 kb, respectively. The results indicate that this standard fingerpriting patterns of DNA hybridization with 4.7 kb probe can be further used for genotyping clinical isolates of Actinobacillus 8ctinomycetemcomitansand its relevance with periodontal disease activity.

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

• Microbiology and Biotechnology Letters
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• v.24 no.2
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• pp.242-246
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• 1996

Hatomarubigins inhibited the growth of various cancer cell lines including multidrug-resistance cells. Hatomarubigins were found to potentiate the colchicine- and vinblastine-induced cytotoxicity against KB-C2 cell, but not the adriamycin-induced cytotoxicity against KB-C2 cells. Hatomarubigins didn't affect the sensitive KB cells. These results suggest that hatomarubigins are specific potentiators of colchicine. Among four hatomarubigins, hatomarubigin A sho- wed the highest synergestic effect on colchine-induced cytotoxicity. Similar effect of hatomarubigin A was found against V79/ADM cells.

• Korean Journal of Poultry Science
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• v.46 no.1
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• pp.17-24
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• 2019

Chickens have been considered as well-defined animal bioreactor. The optimized ovalbumin promoter is essential for recombinant protein production in transgenic chicken. Here we try to compare the activity and identify the effect of estrogen on ovalbumin promoter according to each promoter length with estrogen response element (ERE) existence. We cloned two (2.8 and 5.5 kb) ovalbumin promoters that the 5.5 kb contained the ERE but the 2.8 kb did not, and these two promoters were cloned to pGL4.11 vector. Additionally, we constructed another pGL4.11 vector containing of the 4.4 kb (with ERE) ovalbumin promoter deleted with 1 kb between ERE region and the 2.8 kb promoter. For reporter assay, HeLa, MES-SA, LMH/2A, and cEF cells were transfected with all the pGL4.11 vectors. The comparative analysis showed that the mutated 4.4 kb promoter has more potent activity than the 2.8 and 5.5 kb promoters in HeLa, MES-SA, and LMH/2A cells. However, there is no significant difference in cEFs. Also, these cells transfected with the mutated 4.4 kb promoter were treated with the $17{\beta}$-estradiol (0~3,000 nM) and HeLa, MES-SA, and LMH/2A cells showed estrogen responsibilities, but cEFs did not. Besides, the mutated 4.4 kb promoter has still higher activity than the 2.8 and 5.5 kb promoter, and there is no transcriptional induction effect in 2.8 kb promoter at 500 nM estrogen that is blood concentration of laying hens. Hence our study strongly suggested that the mutated 4.4 kb promoter is considered as one of the most efficient length for generating transgenic chicken.

• Microbiology and Biotechnology Letters
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• v.21 no.5
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• pp.473-477
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• 1993

Both glycerol-phosphate acyltransferase (GPAT) and 7.2 kb mRNAs were present at the highest level in liver. Glycerol-phosphate acyltransferase and 7.2 kb mRNA levels increased dramatically when fasted mice were refed a high carbohydrate diet. In mature 3T3-L1 adipocytes, insulin increased both glycerol-phosphate acyltransferase and 7.2kb mRNA levels 2.6 to 3-fold while dibutyryl cAMP decreased mRNA levels by 50% and 80%, respectively. These results indicate positive regulation by insulin and negative regulation by dibutyryl cAMP of both glycerol-phosphate acyltransferase and 7.2 kb mRNA.

• IMMUNE NETWORK
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• v.8 no.4
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• pp.137-142
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• 2008

Background: CD11c, also known as integrin alpha x, is one of the optimum markers of dendritic cells. However, the regulation of the CD11c expression in mouse has not been identified yet. In this study, in order to analyze the regulation of CD11c expression, the promoter of CD11c was cloned and characterized. Methods: To identify the promoter portion, various sizes of what are considered to be CD11c promoter fragments was amplified by polymerase chain reaction (PCR), using mouse genomic DNA as a template. After sequence was obtained, these fragments were transfected into various cell lines including mouse dendritic cell lines such as JAWSII and DC2.4 and L929 as control cell line.. The promoter activity of three promoter fragments was measured and compared by luciferase activity in the transfected cells. Results: Three clones with size of 1kb, 3kb and 6kb were obtained from mouse genomic DNA. Flow cytometry analysis of JAWSII cells revealed that 52% of the cells expressed CD11c, which was confirmed by RT-PCR analysis. On the contrary, L929 and DC 2.4 cells did not express CD11c. The CD11c+ JAWSII cells were enriched from 52% to 90% with cell sorter. The comparative luciferase activity analyisis demonstrated that the region responsible for tissue specific expression was contained within -3 kb and the clone with size of 3 kb particularly showed higher luciferase activity than 6 kb and 1 kb clones. Conclusion: The CD11c promoter region containing the region responsible for tissue specificity was successfully cloned and -3 kb region showed the highest activity.

• Korean Journal of Microbiology
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• v.47 no.1
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• pp.92-96
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• 2011

Rhodococcus sp. strain DK17 possesses three megaplasmids (380 kb pDK1, 330 kb pDK2, and 750 kb pDK3). The alkylbenzene-degrading genes (akbABCDEF) are present on pDK2 while the phthalate operons which are duplicated are present on both pDK2 (ophA'B'C'R') and pDK3 (ophABCR). DK17 with an optimal temperature of $30^{\circ}C$ showed no growth at $37^{\circ}C$. When transferred to $30^{\circ}C$, however, the $37^{\circ}C$ culture began to grow immediately, indicating that $37^{\circ}C$ is not lethal but stressful for DK17 growth. In addition, when exposed to $37^{\circ}C$ even for a short time, a part of DK17 cells lost the ability to degrade o-xylene (a model compound of alkylbenzenes). When two hundred colonies were randomly selected for colony PCR for pDK2-specific akbC, ophC', or pDK3-specific ophC, a total of 29 colonies were found to have lost at least one of the three genes. PFGE analysis clearly showed that all the mutants have different megaplasmid profiles from that of DK17 wild type, which are divided into five different cases: Type I (10 mutants, pDK2 loss and acquisition of a new ~700 kb plasmid), Type II (9 mutants, pDK2 loss), Type III (8 mutants, pDK3 loss and acquisition of a new ~400 kb plasmid), Type IV (1 mutant, pDK3 loss), and Type V (1 mutant, pDK2 and pDK3 loss and acquisition of the ~400 kb and ~700 kb plasmids). The above results showing that growth temperature changes can induce physical changes in bacterial genomes suggest that environmental changes in habitats including temperature fluctuations affect significantly the evolution of bacteria.

• The Journal of Korean Society of Virology
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• v.29 no.4
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• pp.261-269
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• 1999

The relationship between second messenger cGMP and human cytomegalovirus (HCMV) replication was investigated. First, the intracellular level of cGMP ([cGMP]i) in HCMV-infected cells was measured. The [cGMP]i increased at early times after HCMV infection, reached maximum level at 12 hr and returned to basal level at 24 hr after virus infection, while [cGMP]i in mock-infected cells remained relatively unchanged. Increasing [cGMP]i resulted in enhanced transcription of HCMV major immediate early gene. For early gene expression, cGMP had varying effect. Expression of 1.2 kb RNA decreased and 2.2 kb RNA increased with increasing cGMP, while 2.7 kb RNA gene expression was not affected. HCMV early genes are regulated by immediate early gene, and the effect of cGMP on the regulatory effect of major immediate early gene on early genes was investigated. In the absence of cGMP, major immediate early gene repressed 2.7 kb RNA gene expression, while 1.2 kb RNA and 2.2 kb RNA early genes were not significantly affected. In the presence of $1\;{\mu}M$ cGMP, however, major immediate early gene stimulated the expression of three early genes.

• KSBB Journal
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• v.14 no.1
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• pp.76-81
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• 1999

Characteristics and oxidizing ability of both $NH_4-N$ and$NO^2$-N were examined for the strains isolated from wastewater treatment facilities and from natural systems by using Winogradsky columns. In case of $NH_4$-N, the most efficient strain was Nitrosomonas KB1 isolated from wastewater treatment facility of K corporation and in case of $NO_2$-N, it was Nitrobacter KB2 from the same site as Nitrosomonas KB1. For Nitrosomonas KB1, 91% of $NH_4$-N was oxidized after 4 days of cultivation. Optimal growth temperature and initial pH of Nitrosomonas KB1 were $28^{\circ}C$ and 7, respectively. In comparison to oxidizing rates with changing initial concentration of $NH_4$-N, the ammonia oxidizing rate was increased up to 6.7 mg/day for the initial $NO_2$-N concentrations for the region lower than 100 mg $NH_4-N/L$, but it was gradually reduced for the region higher than 100 mg $NH_4-N/L$. For Nitrobacter KB2 90% of $NO_2$-N was removed after culturing for 4 days. Optimal growth temperature and initial pH of Nitrobacter KB2 was $28^{\circ}C$ and 7, respectively. And the nitrite oxidizing rate was increased in proportion to the initial concentrations of $NO_2$-N up to 200 mg/$\ell$, and it was maintained almost 4.2 mg/day irrespective of initial $NO_2$-N higher than 200 mg/L.

• Korean Journal of Microbiology
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• v.30 no.3
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• pp.218-224
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• 1992

Recombinant plasmid shuttle vectors were constructed for genetic studies on the oxidation of carbon monoxide by carboxydobacteria. Two vectors. pYK322 (7.2 kb, Ap'. Tc') and pYK 324 (7.2 kb, Ap', Tc'), were constructed using pBR322 and pYK100. a small plasmid in Pseudomonas carbo,xydovorans. Four plasmids. pYK2IO (5.2 kb. Cm'), pYK220 (5.2 kb, Cmr), pYK230 (5.2 kb, Cm'), and pYK232 (5.2 kb. Cm'), were constructed using pACYC184 and pYK100. Transformation of several carboxydobacteria with pYK322 and pYK220 was round to be efficient when the cells were transformed by the methoti of Bagdasarian and Timmis (Curr. Top. Microbiol. Immunol. 96:47-67. 1982) with several modifications; cells growing on 0.2% succinate were harvested at the mid-exponential phase. 10 mM RbCl in transformation solution was substituted with 100 mM KCI. cclls in transformation solution were incubated for 12 h at 4'C before addition of DNA and heat shock was carried out for 3 min at 45$^{\circ}$C. Plasmid vectors used for transformation, however. were not detected from antibiotics-resistant transformants, suggesting that the vectors may be integrated into the chromosomal DNA.