• Journal of Life Science
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• v.12 no.3
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• pp.340-348
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• 2002

Equisetum arvense L. and Lactuca dentata Makino. var. Flaviflora Makino. of samples relatively showed anticancer effects on MCF-7 mammary gland adenocarcinoma cell. The most active plant among the samples was Capsicum annuum L. var angulosum Mill. We studied that MCF-7 cells were changing chromosomal morphology and apoptosis on these samples. Capsicum annuum L. var. angulosum Mill. of samples relatively showed good anticancer effects. The cells became vague after 2 days and then destroyed. The supernatant of the cells including medium was measured by UV absorbance. The results showed that Capsicum annuum L. var. angulosum Mill also exerted high level. We also used electrophoresis in order to observe apoptic characterization of DNA fragmentation. The cells treated with Capsicum annuum L. var. angulosum Mill showed the apoptotic characterization. The chromosome of the cells were observed on those samples. The cells treated with Capsicum annuum L. var. angulosum Mill among them were shown the fastest changes. The cells were aggregated and destroyed by treatment with some edible plants. Especially, the case of Capsicum annuum L. var. angulosum Mill, it led MCF-7 cell to apoptosis faster than others. And we can observe chromosomal changes and dispersion by PI staining. These results showed that each sample exerted anticancer effects on MCF-7 cells. Especially Capsicum annuum L. var. angulosum Miff exerted significant anticancer effects.

• Archives of Pharmacal Research
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• v.21 no.3
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• pp.310-314
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• 1998

DW-116 is a new fluoroquinolone antimicrobial agent with a broad spectrum. In order to elucidate the resistance mechanism to DW-116 in Acinetobacter spp. bacteria, total chromosomal DNA was isolated from 10 strains of Acinetobacter spp. resistant to DW-116. Quinolone resistance determinant region (QRDR) of DNA gyrase gene was amplified by PCR. The 345 bp nucleotide fragment yielded was inserted into pKF 3 which was used as the vector. Comparisons of the DNA sequences of 8 strains with that of the wild type strain revealed a Ser-83 to Leu mutation in mutants and all ten strains contained one silent mutation$(T{\rightarrow}G)$in QRDR. From Acinetobacter MB4-8 strain, DNA gyrase was isolated and purified, through novobiocin-sepharose, heparin-sepharose affinity column chromatography. The enzyme was composed of two subunits and the molecular mass of subunits A and B were 75.6 and 51.9 kDa, respectively. The supercoiling activity of the reconstituted DNA gyrase composed of subunit A from Acinetobacter MB4-8 and subunit B from E. coli was not inhibited by $128{\mu}\textrm{g}$ml of ciprofloxacin. It might be said that one of the resistance mechanisms to DW-116 in Acinetohacter MB4-8 was subunit A alteration of DNA gyrase.

• Korean Journal of Plant Resources
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• v.20 no.5
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• pp.446-450
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• 2007

Tiarella polyphylla D. Don(Saxifragaceae) is a perennial herb and distributed in China, Japan, Taiwan and Korea. Especially, it only grows in Ulleung island of Korea. It has been using for asthma, bruise and audition troubles with main components of some Triterpenoids and seven oleanolic Saponins. There is only known its chromosomal number rarely and cytogenetic study was not done. From this study, karyotype analysis and chromosomal localization of 5S and 45S rDNAs using bicolor-FISH(fluorescence in situ hybridization) were carried out. The somatic metaphase chromosome number was 2n=2x=14 and the size of chromosomes ranged $1.66{\sim}3.50{\mu}m$. The chromosome complement consisted of four pairs of submetacentrics(chromosomes 1, 2, 3 and 6), two pairs of subtelocentrics(chromosomes 5 and 7) and one pair of telocentrics(chromosome 4). We also observed NOR(nucleolus organizer region) on the chromosome 4. In bicolor-FISH, one pair of 55 and 45S rDNA sites was detected on the centromeric region of chromosome 3 and short arm of chromosome 4, respectively. Bicolor FISH was very useful tool for the localization and identification of rDNAs on the chromosomes in Tiarella polyphylla.

• Microbiology and Biotechnology Letters
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• v.19 no.1
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• pp.21-24
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• 1991

The promoter isolated from an alkali-tolerant Hwillus sp. chromosomal DNA was subcluned. The activity of promoter in B, subtilis and alkali-tolerant Bacillus sp. began to increase at the early stage. of spore formation. In the presence of 1% glucose, the promotcr activity repressed and was recovered by ;tddition of c-GMP in the medium.

• Molecules and Cells
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• v.37 no.2
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• pp.95-99
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• 2014

Cancer is unique amongst human diseases in that its cellular manifestations arise and evolve through the acquisition of somatic alterations in the genome. In particular, instability in the number and structure of chromosomes is a near-universal feature of the genomic alterations associated with epithelial cancers, and is triggered by the inactivation of tumour suppressor mechanisms that preserve chromosome integrity in normal cells. The nature of these mechanisms, and how their inactivation promotes carcinogenesis, remains enigmatic. I will review recent work from our laboratory on the tumour suppressor BRCA2 that addresses these issues, focusing on new insights into cancer pathogenesis and therapy that are emerging from improved understanding of the molecular basis of chromosomal instability in BRCA2-deficient cancer cells.

• Biomedical Science Letters
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• v.25 no.3
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• pp.275-281
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• 2019

This study was performed to characterize the chromosomal metallo-${\beta}$-lactamases (MBLs) of Chryseobacterium indologenes isolated from Korea and to propose a clustering method of IND MBLs based on their amino acid similarities. Chromosomal MBL genes were amplified by PCR from 31 clinical isolates of E. indologenes. Nucleotide sequencing was performed by the dideoxy chain termination method using these PCR products. Antimicrobial susceptibilities were determined by the agar dilution method. PCR experiments showed that all 31 E. indologenes isolates contained all $bla_{IND}$ genes. DNA sequence analysis revealed that E. indologenes isolates possessed ten types of $bla_{IND}$ gene, including seven novel variants ($bla_{IND-8}$ to $bla_{IND-14}$). The most common combination of MBL was IND-2 (n = 18). Minimum inhibitory concentrations of imipenem and meropenem for the isolates harboring novel IND MBLs were ${\geq}16{\mu}g/mL$. IND MBLs were grouped in three clusters, based on amino acid similarities.

• Journal of Life Science
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• v.15 no.5 s.72
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• pp.734-738
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• 2005

A gene coding for xylanase from alkali-tolerant Bacillus alcalophilus AX2000 was cloned into Escherichia coli $DH5\alpha$ using pUC19. Among 2,000 transformants, one transformant showed clear zone on the detection agar plate containing oat-spells xylan. Its recombinant plasmid, named pXTY99, was found to carry 7.0 kb insert DNA fragment. When the nucleotide sequence of the cloned xylanase gene (xynT) was determined, xynT gene was found to consist of 1,020 base-pair open reading frame coding for a poly-peptide of 340 amino acids with a deduced molecular weight of 40 kDa. The coding sequence was preceded by a putative ribosome binding site, and the transcription initiation signals. The deduced amino acid sequence of xylanase is similar to those of the xylanases from Bacillus sp. Nl37 and B. stearothermophilus 21 with $61\%$ and $59\%$ identical residues, respectively.

• Journal of Microbiology and Biotechnology
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• v.7 no.4
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• pp.229-236
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• 1997

A gene, $phbC_{2.4.1}$ encoding poly-3-hydroxybutyric acid (PHB) synthase of Rhodobacter sphaeroides 2.4.1 was cloned by employing heterologous expression in Escherichia coli. R. sphaeroides chromosomal DNA partially digested with MboI was cloned in pUC19 followed by mobilization into E. coli harbouring $phbA,B_{AC}$ in pRK415, which code for ${\beta}$-ketothiolase and acetoacetyl CoA reductase of Alcaligenes eutrophus, respectively. Two E. coli clones carrying R. sphaeroides chromosomal fragment of $phbC_{2.4.1}$ in pUC19 were selected from ca. 10,000 colonies. The PHB-producing colonies had an opaque white appearance due to the intracellular accumulation of PHB. The structure of PHB produced by the recombinant E. coli as well as from R. sphaeroides 2.4.1 was confirmed by [$H^{+}$]-nuclear magnetic resonance (NMR) spectroscopy. Restriction analysis of the two pUC19 clones revealed that one insert DNA fragment is contained as a part of the other cloned fragment. An open reading frame of 601 amino acids of $phbC_{2.4.1}$ with approximate M.W. of 66 kDa was found from nucleotide sequence determination of the 2.8-kb SaiI-PstI restriction endonuclease fragment which had been narrowed down to support PHB synthesis through heterologous expression in the E. coli harbouring $phbA,B_{AC}$. The promoter (s) of the $phbC_{2.4.1}$ were localized within a 340-bp DNA region upstream of the $phbC_{2.4.1}$ start codon according to heterologous expression analysis.

• Applied Biological Chemistry
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• v.45 no.4
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• pp.190-194
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• 2002

Expression of Bacillus subtilis glutamyl-tRNA synthetase (GluRS) in Escherichia coli is lethal for the host, probably because this enzyme misaminoacylates ${tRNA_l}^{Gln}$ with glutamate in vivo. In order to overexpress B. subtilis GluRS, encoded by the gltX gene, in E. coli, this gene was amplified from B. subtilis 168 chromosomal DNA using PCR method and the entire coding region was cloned into a pET11a expression vector so that it was expressed under the control or the T7 Promoter. The resulting recombinant pEBER plasmid was transformed into E. coli Novablue (DE3) bearing the T7 RNA polymerase gene for expression. After IPTG treatment, the overproduced enzyme was purified using ammonium sulfate fractionation, Source Q anion exchange chromatography, Superdex-200 gel filtration, and Mono Q anion exchange chromatography. The purified enzyme yielded 18-fold increase in specific activity over the crude cell extract and its molecular weight was approximately 55 kDa on SDS-PAGE.

• Applied Biological Chemistry
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• v.38 no.1
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• pp.7-12
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• 1995

The purpose of this study was the development of promoter for the lacZ' gene. Two heterologous promoter I and II of lacZ' gene were isolated from chromosomal DNA Bam HI fragment of yeast. The size of the promoter I was estimated to be 2.5 kb and ${\beta}-galactosidase$ activity was 124.6 U/mg protein, and the size of the promoter II was 4.0 kb and its ${\beta}-galactosidase$ activity was 168.8 U/mg protein, respectively. The stability of the recombinant YEp plasmid in the transformant was from 52.7 to 67.4% at minimal medium. YIp plasmid was constructed from YEp plasmid, and expressed both in E. coli and yeast. The promoter I aid II iso-lated from yeast chromosomal DNA can be used for promoter of plasmid YEp and YIp.