• Microbiology and Biotechnology Letters
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• v.35 no.2
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• pp.104-111
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• 2007

In this study, aniline dioxygenase genes responsible for initial catabolism of aniline in Burkholderia sp. HY1 and Delftia sp. HY99 were cloned and the amino acid sequences were comparatively analyzed, which already have been reported as bacteria utilizing aniline as a sole source of carbon and nitrogen, B. sp. HY1 was found to have at least a plasmid, and the plasmld-cured strain, B. sp. HY1-PC obtained using mitomycin C was tested with wild type strain to investigate whether the former maintained the degradability for aniline. This proved that the aniline oxygenase gene from B. sp. HY1 was located in chromosomal DNA, not in plasmid DNA. Aniline dioxygenase small subunits from B. sp. HY1 and D. sp. HY99 were found, based on 146 amino acids, to share 79% similarity. Notably, ado2 genes from B. sp. HY1 and D. sp. HY99 which were found to be terminal dioxygenase of aniline dioxygenase small subunit showed 99% similarity in the deduced amino acid sequences with tdnA2 of Frateuria sp. ANA-18 and danA2 of D. sp. AN3, respectively. Besides, enzyme assay and amino acid sequence analysis of catechol dioxygenase supported the previous report that B. sp. HY1 might occupy ortho-cleavage pathway using catechol 1,2-dioxygenase, while D. sp. HY99 might occupy catechol 2,3-dioxygenase for meta-cleavage pathway.

• Journal of the korean academy of Pediatric Dentistry
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• v.33 no.2
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• pp.304-310
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• 2006

Prevotella intermedia is one of the most frequently implicated pathogens in human periodontal disease and has a requirement for hemin for growth. This study has identified a hemin-binding P. intermedia protein by expression of a P. intermedia genomic library in Escherichia coli, a bacterium which does not require or transport exogenous hemin. The genomic library of P. intermedia was constructed into plasmid pUC18, transformed into Escherichia coli strain $DH5{\alpha}$, and screened for recombinant clones using heminbinding activity by plating onto hemin-containing agar. Approximately 5,000 recombinant E. coli colonies were screened onto LB-amp-hemin agar, single clone(pHem1) was exhibited a clearly pigmented phonotype. The 2.5kb insert DNA of pHem1 was determined by restriction enzyme mapping. Southern blot analysis of BamHI, BglII, EcoRI, HindIII and PstI-digested P. intermedia DNA indicated that single copy of the gene was present in the genome. Northern blot analysis revealed that the size of transcript was approximately 1.8 kb. The cloned gene contained a single ORF, consisting of approximately 850-residue amino acids. A BLAST search of the Institute for Genomic Research genes with similar nucleotide sequence revealed no significant similarity It needs further investigation to clarify the mechanisms of heme uptake in P. intermedia.

• Biomedical Science Letters
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• v.5 no.2
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• pp.135-145
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• 1999

A total of 45 Staphylococcus aureus strains from clinical samples were tested for the biochemical test and antibiotic susceptibility test. Forty-five S. aureus strains were subjected to the molecular epidemiological study by susceptiblity test, antibiogram, bacteriophage typing, polymerase chain reaction and mec-associated hypervariable region gene in order to detect of mecA gene which was one of the structural gene related to antibiotic resistant expression factors. Three of 15 mecA-negative S. aureus isolates were classified as oxacillin resistant despite borderline minimal inhibitory concentration values. Methicillin susceptiblities were completely consistent with PCR results for these strains. On the other hand, 4 of 30 mecA-positive isolates yielded results in the oxacillin and methicillin susceptibility tests which were discrepant from those of PCR analysis. Except for SA6, the methicillin resistant S. aureus strains tested were highly resistant to penicillin, oxacillin, gentamicin, and chloramphenicol. In the phage typing, 27 strains were typable. The Iytic group III was as many as 12 strains, and 7 of 12 were 75/83A/84 type. In the PCR of specific mecA gene probe with chromosomal DNA of 30 methicillin resistant S. aureus, the amplified DNA band of 533 bp was confirmed in 30 strains and not in methicillin sensitive S. aureus. The single amplified band of hypervariable region related to mec was investigated in all of 30 methicillin resistant S. aureus, but in methicillin sensitive S. aureus it was amplified. The size of PCR products was between 200 bp and 600 Up. Four units was directly repeated.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.3
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• pp.363-368
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• 1999

This study was to confirm whether the vitrification method using EFS40 freezing solution has detrimental effect on the cytoskeleton and chromosome constitution of the immature mouse oocytes by indirect immunocytochemistry and chromosome analysis. Immature mouse oocytes were vitrified using EFS40 (40% EG, 18% ficoll, 0.5 M sucrose diluted in M2 medium), thawed and then survived oocytes were in vitro matured for 16 hr. When the microtubule morphology and micro filament distribution in vitrified-thawed immature mouse oocytes were examined, normal percentage of two cytoskeleton in vitrified group (93.9 and 100.0%) was not significantly different from that in control (100.0 and 100.0%) and exposed group (94.4 and 100.0%). The rate of oocytes containing a normal chromosome number in vitrified group was 65.8%, this result was not significantly different from that in control (79.6%) and exposed group (69.0%). These results indicated that exposure to cryoprotectant or freezing has not effect on the alteration of cytoskeleton morphology and the chromosome constitution of mouse oocytes and that our vitrification methods using EFS40 freezing solution was suitable for the cryopreservation of immature mouse oocytes.

• Tuberculosis and Respiratory Diseases
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• v.42 no.4
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• pp.513-521
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• 1995

Background: Nucleolar organizer regions(NORs) are chromosomal segments encoding for ribosomal RNA and associated with argyrophilic nonhistone protein. Ribosomal RNA genes ultimately direct ribosome and protein synthesis, and it has been suggested the numbers of NORs detected in the cell may reflect nuclear and cellular activity. This study was performed to evaluate the applicability of AgNORs to the diagnosis of squamous cell carcinoma of the lung. Method: The one step silver methods(AgNORs) was used to stain NORs in the routinely processed, formalin fixed, paraffin embedded sections of 36 cases of squamous cell carcinoma of the lung obtained by surgical resection of primary tumor. In each specimen, 100 tumor cells and 100 normal cells adjacent to the tumor chosen at random were examined under an oil immersion lens at a magnification of ${\times}1000$. The mean number of AgNORs per nucleus was calculated for each specimen. Results: The mean number of AgNORs per nucleus(mAgNORs) of normal bronchial epithelium and squamous cell carcinoma of the lung was $1.74{\pm}0.25$ and $4.05{\pm}0.80$, respectively. The difference of mAgNOR between normal and tumor tissue was statistically significant(p<0.001). There was no statistical difference among tumors of different stages. The difference of mAgNOR between normal and tumor tissue was statistically significant in each TNM stage(p<0.05). Conclusion: Mean AgNOR count may be used as a useful marker for the differential diagnosis of benignancy and malignancy, and proliferative activity of the cell in squamous cell carcinoma of the lung. But there was no statistical difference in mean AgNOR count among tumors of different surgical stages. Further studies for the application of mAgNORs to the diagnosis of other histologic types and cytologic specimens of the lung cancer are needed.

• Journal of Radiation Protection and Research
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• v.28 no.2
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• pp.117-125
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• 2003

It was reported that radiopharamaceuticals induced radiation adaptive response (RAR) in patients undergoing nuclear medicine imaging studies. Individual variations of RAR were not studied well. The purpose of this study was to evaluate individual variation of RAR in patients undergoing nuclear medicine imaging studies. Peripheral lymphocytes were collected from 23 patients undergoing $^{99m}Tc-diethylenetriamine$ pentaacetic acid $(^{99m}Tc-DTPA)$ renal scintigraphy, 18 patients undergoing $^{99m}Tc-methylene$ diphosphonate $(^{99m}Tc-MDP)$ bone scintigraphy and 21 patients undergoing $^{99m}Tc-tetrofosmin\;(^{99m}Tc-TF)$ scintigraphy were collected before and 4 hours after injection of radiopharmaceuticals. The lymphocytes were exposed to challenge dose of 2 Gy gamma rays using a cell irradiator. Numbers of ring-form (R) and dicentric (D) chromosomes were counted under the light microscope. and used to calculate the frequency of chromosomal aberration [Ydr=(D+R)/total number of counted lymphocytes]. Adaptation index (k) was defined 3s ratio of Ydr in conditioned lymphocytes over Ydr in unconditioned lymphocytes. Coefficients of variance of k in $^{99m}Tc-DTPA,\;^{99m}Tc-MDP\;and\;^{99m}Tc-TF$ were 35%, 34% and 21%, respectively k was not dependent upon age, sex, and underlying diseases. There was a wide variation of RAR induced by radiopharmaceuticals among patients undergoing nuclear medicine procedures. It remains to be determined for causes of such variation.

• Food Science and Preservation
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• v.24 no.1
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• pp.100-106
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• 2017

This study was conducted to evaluate the genotoxicity of balanced nutritional formular for patients containing various ingredients after gamma irradiation at 4 kGy. Since viable bacteria were not observed within the detection limit of 1 log CFU/g, a dose of 4 kGy was appropriate for the pasteurization of the formular. In a bacterial reverse mutation assay, both hot water and methanol extracts of the formular exhibited dose-independent responses, which was similar to those obtained from that of the negative control (distilled water or dimethyl sulfoxide). In a chromosomal aberration test using lung fibroblast cells of Chinese hamster, the numbers of normal chromosomes were comparable to those observed in the negative control, regardless of the treatment dose and metabolic activation system. Furthermore, no significant increases in the frequency of micronucleated polychromatic erythrocytes were observed relative to the control, when mice were fed with the formular at doses up to 2,000 mg/kg body weight. Therefore, the balanced nutritional formular for patients did not exhibit genotoxicity when pasteurization by gamma irradiation at 4 kGy.

• Microbiology and Biotechnology Letters
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• v.30 no.1
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• pp.8-14
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• 2002

Pseudomonas sp. S-47 is capable of degrading 4-chlorobenzoate to produce 5-chloro-2-hydroxymuconic semialdehyde (5C-2HMS) by the enzymes encoding by xylXYZLTE cluster. In this study, the resulting 5C-2HMS was confirmed to be transformed to 5-chloro-2-hydroxymuconic acid (5C-2HMA) by 5C-2HMS dehydrogenase. The xylG gene encoding 5C-2HMS dehydrogenase was cloned from the chromosomal DNA of strain S-47. The nucleotide sequence of xylG showed to be composed of 1,600 base pairs with ATG initiation and TGA termination codons. A deduced amino acid sequence of the 5C-2HMS dehydrogenase (XylG) exhibited 98％, 93％, and 89％ identity with those of the dehydrogenases from P. putida mt-2, P. putida G7, and Pseudomonas sp. CF600, respectively.

• Journal of Plant Biotechnology
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• v.41 no.1
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• pp.19-25
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• 2014

The total production volume has been sharply increased from year 2008 in Gyeongnam province, South Korea by the policy of preservation and promotion of indigenous balloon flower germ lines. In an attempt to assist the Gyeongnam province's policy, in this study, we tried to establish a technique to differentiate the indigenous balloon flower germ lines with those collected within South Korea and China. Our preliminary results indicated that RAPD analyses with five different primers exhibited high frequency of polymorphic DNA bands up to 76.9% and phylogenetic tree indicated that some of the indigenous lines can be easily differentiated with others. However, it was suggested that more advanced techniques such as single nucleotide polymorphic markers need to be developed in particular, by using extra-chromosomal DNA.

• Journal of Life Science
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• v.17 no.5 s.85
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• pp.723-727
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• 2007

Resistance to metronidazole in Helicobacter pylori results from inactivation of rdxA and frxA, the chromosomal genes for a nitroreductase that normally converts metronidazole from prodrug to bactericidal agent. Two types of metronidazole susceptible strains had been found distinguishable by their apparent levels of frxA expression. Most common in the populations we had studied were strains that required only rdxA inactivation to become resistant to moderate levels of metronidazole(type I strains). The second strain type required inactivation of both frxA and rdxA to become resistance to metronidazole(type II strains): this was linked to a relatively high level of frxA gene transcription in the type II strains. The fdxA gene regulated fdxA as well as rdxA gene. Thus, to study the function of fdxA as a regulatory gene we constructed a null mutant of fdxA in H. pylori genome and identified over-and under-expressed proteins by fdxA using two-dimensional(2-D) electrophoresis and MALDI-TOP-MS. There were four over-expressed proteins in fdxA mutant; nifU-like protein(HP0221), frxA(HP0642), nonheme ferritin(HP0653), and hypothetical protein(HP0902). Three under-expressed proteins were also identified in fdxA mutant, including 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase (HP0089), (3R)-hydroxymyristoyl ACP dehydratase(HP1376), and thioredoxin(HP1458).