• Journal of Microbiology and Biotechnology
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• v.33 no.2
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• pp.211-218
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• 2023

A cryptic plasmid (pTH32) was characterized from Tetragenococcus halophilus 32, an isolate from jeotgal, Korean traditional fermented seafood. pTH32 is 3,198 bp in size with G+C content of 35.84%, and contains 4 open reading frames (ORFs). orf1 and orf2 are 456 bp and 273 bp in size, respectively, and their translation products showed 65.16% and 69.35% similarities with RepB family plasmid replication initiators, respectively, suggesting the rolling-circle replication (RCR) mode of pTH32. orf3 and orf4 encodes putative hypothetical protein of 186 and 76 amino acids, respectively. A novel Tetragenococcus-Escherichia coli shuttle vector, pMJ32E (7.3 kb, Em^r), was constructed by ligation of pTH32 with pBluescript II KS(+) and an erythromycin resistance gene (ErmC). pMJ32E successfully replicated in Enterococcus faecalis 29212 and T. halophilus 31 but not in other LAB species. A pepA gene, encoding aminopeptidase A (PepA) from T. halophilus CY54, was successfully expressed in T. halophilus 31 using pMJ32E. The transformant (TF) showed higher PepA activity (49.8 U/mg protein) than T. halophilus 31 cell (control). When T. halophilus 31 TF was subculturd in MRS broth without antibiotic at 48 h intervals, 53.8% of cells retained pMJ32E after 96 h, and only 2.4% of cells retained pMJ32E after 14 days, supporting the RCR mode of pTH32. pMJ32E could be useful for the genetic engineering of Tetragenococcus and Enterococcus species.

• Journal of Microbiology
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• v.41 no.2
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• pp.89-94
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• 2003

Bacterial strain No. P08 isolated from wastewater at the Cheongju industrial complex was found to be capable of degrading 4-chlorobenzoate under aerobic condition. P08 was identified as Comamonas sp. from its cellular fatty acid composition and 16S rDNA sequence. The fcb genes, responsible for the hydrolytic dechlorination of 4-chlorobenzoate, were cloned from the plasmid pJJl of Comamonas sp. P08. The fcb gene cluster of comamonas sp. PO8 was organized in the order fcbB-fcbA-fcbTl-fcbT2-fcbT3-fcbC. This organization of the fcb genes was very similar to that of the fcb genes carried on the chromosomal DNA of pseudomonas sp. DJ-12. However, it differed from the fcbA-fcbB -fcbC ordering of Arthrobacter sp. SU. The nucleotide sequences of the fcbABC genes of strain P08 showed 98% and 53% identities to those of Pseudomonas sp. DJ-12 and Arthrobacter sp. SU, respectively. This suggests that the fcb genes might have been derived from Pseudomonas sp. DJ-12 to form plasmid pJSl in Comamonas sp. P08, or that the fcb genes in strain DJ-12 were transposed from Comamonas sp. P08 plasmid.

• Korean Journal of Microbiology
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• v.27 no.3
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• pp.181-187
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• 1989

Plasmid DNA was isolated from Lactobacillus casei SW-M1($Lac^{+}$strain). The curing frequencies of pPLac plasmid from L. casei SW-M1 showed 43% for acriflavin treatment and 53% for ethidium bromide treatment after 3 times transfer. On the charaterization of pPLac plasmid, it was found that the plasmid contained gene encoding phospho-$\beta$-galactosidase for lactose utilization. Lactose-PTS(phosphotransferase system)was involved in membrane transport system in $Lac^{+}$ strain. Induction of phospho-$\beta$-galactosidase was specially effective by galactose, lower effect with lactose and glucose but not by IPTG(isopropyl-$\beta$-D-thiogalactoside). This result showed that induction of phospho-$\beta$-galactosidase by IPTG did not appeared. The catabolite repression of phospho-$\beta$-galactosidase synthesis by glucose was not found in L. casei.

• Environmental Mutagens and Carcinogens
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• v.25 no.1
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• pp.40-46
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• 2005

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) displays high toxicity in animals and has been implicated in human carcinogenesis. Although the mechanism of carcinogenesis by TCDD is unclear, it is considered to be a non-genotoxic compound and tumor promoter. In our experiment, we investigated the effects of TCDD on gene expression in mouse skin carcinogenesis. We used cDNA microarray to detect the differential gene expression in tumors induced in hairless mouse skin by MNNG plus TCDD protocol. We found that erb-2, c-ets2 and p27$^{kip1}$ were significantly up-regulated, but TNFR2, AKT-l, integrin $\beta$l, maspin, IGF-l, c-raf-l, Rb were significantly down-regulated, in tumor region, respectively. We also found that the expression of 53 genes involved in cen cycle, signal transduction, apoptosis, adhesion molecule, angiogenesis, and invasion, were changed two fold more, in tumor surrounding region. These data suggest that TCDD alters the expression of a large array of genes involved in apoptosis, cytokine production and angiogenesis in mouse skin carcinogenesis.

• Proceedings of the Zoological Society Korea Conference
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• 1998.10b
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• pp.310.1-310
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• 1998

No Abstract, See Full Text

• Proceedings of the Zoological Society Korea Conference
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• 1999.10b
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• pp.250.1-250
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• 1999

No Abstract, See Full Text

• 대한예방의학회:학술대회논문집
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• 1999.10a
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• pp.1-15
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• 1999

In a case-control study to evaluate the factors involved in the development of hepatocellular carcinoma, polymorphisms of the p53 gene were compared in 68 cases mostly infected with hepatitis C virus (HCV) and 68 controls matched for sex and age: DNA from peripheral blood leukocytes was analyzed by the polymerase chain reaction-single strand conformation polymorphism method and direct sequencing. Polymorphisms analyzed were those in exon 4 (CCC vs. CGC, Pro vs. Arg at codon 72, Al allele vs. A2 allele), intron 2 (C vs. G at nucleotide 38, Al vs. A2), intron 3 (C vs. A at nucleotide 65, Al vs. A2; absence and presence of 16 base pair repeat at nucleotides 24 to 39, Al vs. A2), intron 6 (A vs. G at nucleotide 62, Al vs. A2) and intron 7 (C and T vs. T and G at nucleotides 72 and 92, Al vs. A2). A significantly higher frequency of the allele for CCC (Pro, Al) at codon 72 of exon 4 was found in cases (39%) than in controls (26%) (p<0.05). Highly significant linkage of the polymorphisms in exon 4, intron 2, intron 3 and intron 7, and between the intron 3-16 bp duplication and polymorphism in intron 6 also was found. Matched Fair analysis showed significantly higher frequencies of certain haplotypes (1-1-1-1-2-2 or 1-1-2-1-2-1 for exon 4, intron 2, intron 3, the intron 3-16 bp duplication, intron 6 and intron 7) in cases than in controls (p=0.014, OR=2.27, 95% CI= 1.08-5.12). No preference of specific p53 polymorphisms for specific HCV genotype was detected. These findings suggest that in hepatocarcinogenesis mainly due to HCV infection, genetic factors may be involved and that genetic markers can serve as predictors of a high-risk group for hepatocarcinogenesis.

• The Korean Journal of Food And Nutrition
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• v.6 no.1
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• pp.37-46
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• 1993

Brain, heart, liver, lung, kidney and thymus etc. 12 organs were removed and homogenized from Dawley-Sprague rats after suffocation. After fractionation of the tissue cytosols, enzymatic activities of the key enzymes in metabolic inositol phosphates cycle, PLC, IPSK and Ins(1,4,5) P35-phosphatase, were measured respectively. Hybridoma monoclones producing anti-lP3K murine monoclonal antibodies were obtained by the fusion of SP2/Ag 0-14 and spleen cells of mouse immunized with purified 53KDa IPSK, screening and cloning procedures. 18 cloned hybridoma cells were obtained, background due to nonspecific binding was very low with 10 clones. These Abs were purified from ascitic fluids by using affi-gel 15, and determined subtype of Abs. When immunoreactivities for rat tissues IP3K were exercised by adding the mixed Abs of 19Gl and 19G2b, they showed an overall similarity with noncompetitive inhibition. Brain tissue has high sensitivity for anti-lP3K Ab, whereas heart tissue has very low activity. In kinetic parameters Km value was 1.58 mM and Vmx value was 5.41umol/min/ml, respectively Only one form of 40 KDa IPSK was detected in heart tissues, however rat brain contains at least three immunologically distinct IP3K (53, 51 and 40 KDa) in western blot analysis. Of them 53 KDa protein was major enzyme in enzymatic activity. Northern blot analysis with 32P-labeled CDNA probe which encodes 1.8 Kb IPSK gene was performed. These results suggest that IPSK are regulated at transcriptional level during rat tissue development.

• Korean Journal of Clinical Laboratory Science
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• v.38 no.1
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• pp.45-53
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• 2006

Anaerobic black-pigmented bacteria have been implicated in the endodontic infections. This group of microorganisms includes Porphyromonas endodontalis, P. gingivalis, Prevotella intermedia, and Prevotella nigrescens. The organisms display a wide variety of virulence factors that may be pertinent to acute endodontic infections. The aim of this study was to identify P. endodontalis, P. gingivalis, P. intermedia, and P. nigrescens by using the special potency disk test, filter paper spot test, 16S rRNA gene-directed PCR, and API 32A system. Microbial samples were collected from root canals of 33 intact teeth with necrotic pulp and apical periodontitis. Conventional laboratory methods were used to identify the strains of anaerobic black pigmented bacteria. Eighteen out of 33 samples were positive for the growth of black-pigmented bacrteria. Five colonies were cultured from each pure cultured colony from Brucella agar plates. Seventy seven colonies were positive for the growth of black-pigmented bacteria. Thirty three out of 77(42.8%) were identifed as P. nigrescens, 10 out of 77(13%)were P. gingivalis, 6 out of 77(7.8%) were P. endodontalis, 10 out of 77(13%) were P. intermedia. On the contrary the reference strains of P. nigrescens, experimental strains of P. nigrescens were susceptible to kanamycin in the special potency disk test. We concluded that after rapid presumptive identification methods, such as the special potency disk test and filter paper spot test were done, 16S rRNA gene PCR and API 32A test would be accurate detection methods for black-pigemented bacteria.

• Genomics & Informatics
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• v.8 no.3
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• pp.108-115
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• 2010

Hypertension is the most prevalent disease worldwide and is itself a risk factor for cerebral, cardiac, and renal diseases. The inconsistency of candidate genes suggested by previous genomewide association studies (GWASs) may be due to not only differences in study design and genetic or environmental background but also the difference in the power of analysis between continuous traits and discrete traits. We analyzed 352,228 single nucleotide polymorphisms (SNPs) in 8842 unrelated Koreans obtained from Ansan and Ansung cohorts. We performed a series of GWA analyses using three different phenotype models; young hypertensive cases (278 subjects) versus elderly normotensive controls (680 subjects); the upper 25% (2211 hypertensive cases) versus the lower 25% of the SBP distribution (2211 hypotensive controls); and finally SBP and DBP as continuous traits (8842 subjects). The numbers of young hypertensive cases and elderly normotensive controls were not large enough to achieve genomewide significance. The model comparing the upper 25% subjects to the lower 25% of subjects showed a power that was approximate to that of QTL analysis. Two neighboring SNPs of the ATP2B1 gene, rs17249754 (SBP, p=$2.53^{-10}$; DBP, p=$1.28{\times}10^{-8}$) and rs7136259 (SBP, p=$1.30{\times}10^{-9}$; DBP, p=$6.41{\times}10^{-8}$), were associated with both SBP and DBP. Interestingly, a SNP of the RPL6 gene, rs11066280, revealed a significant genomewide association with SBP in men only (p=$3.85{\times}10^{-8}$), and four SNPs located near the MAN2A1 gene showed a strong association with DBP only in elderly men aged 60-70 years (e.g., rs6421827, p=$4.86{\times}10^{-8}$). However, we did not observe any gene variant attaining genomewide significance consistently in the three phenotype models except for the ATP2B1 gene variants. In general, the association signal with blood pressure was stronger in women than in men. Genes identified in GWASs are expected to open the way for prevention, early diagnosis, and personalized treatment of hypertension.