• Journal of Plant Biotechnology
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• v.31 no.4
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• pp.319-323
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• 2004

Soluble latex protein fraction excreted from Euporbia lathyris laticifer was resolved by 10% SDS-polyacrylamide gel electrophoresis to identify distinctively displayed latex major protein bands including ELp65, ELp55, ELp43, ELp32 and ELp23. Among them, ELp65 was purified by ammonium sulfate precipitation, gel permeation chromatography and ion exchange chromatography. Its N-terminal amino acid sequencing revealed its homology to the leading region of mature peptide of tomato p69a subtilisin-like protease, suggesting a certain role involved in plant defense system. In the analysis of Southern blot hybridization using PCR-amplified tomato p69a probe DNA, E. lathyris genome was suggested to have a gene family consisting of 3-5 gene members putatively encoding subtilisin-like proteases.

• The Korean Journal of Mycology
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• v.17 no.1
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• pp.27-30
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• 1989

F. velutipes Leu 2 gene (${\beta}-isopropylmalate$ dehydrogenase gene) was used for transformation of P. florida leucine requiring auxotrophic mutant P101. Transformation frequency was very low but the transformed colony can grow on minimal medium very slowly. Transformation was identified by Southern hybridization and reverse transformation into E. coli using chromosome DNA isolated from transformed P. florida.

• Journal of fish pathology
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• v.16 no.1
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• pp.61-68
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• 2003

MICs of 8 chemotherapeutic agents against forty streptococcal isolates were determined. These strains were isolated from diseased olive flounder, Paralichthys olivaceus, and showed 2-5 multiple drug resistance against different antibacterial agents including ampicillin, doxycycline (DOXY), erythromycin, florfenicol, flumequine, norfloxacin, oxolinic acid, and oxytetracycline (OTC). In conjugation experiment, we found transferable R plasmids carrying OTC and DOXY resistance determinant in 3 drug resistance strains analyzed. Six out of 40 isolates showed positive signal in colony hybridization with the R plasmid DNA (pST9) as a probe. It suggests that other types R plasmid different from pST9 is also involving in multiple drug resistance of streptococci isolated from olive flounder.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.3
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• pp.423-427
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• 2004

Seventy-six animals including cattle, sheep, horses, 6 species of zoo animals were employed for collection of fresh feces in order to detect verotoxigenic Esherichia coli (VTEC) by safe, quick and sensitive PCR-based molecular methods. Bacterial cell disruption with bead-beating followed by bacterial DNA purification with hydroxyapatide chromatography and gel filtration allowed DNA preparation from animal feces with high recovery and purity. A mountain goat was firstly shown by PCR and sequencing to shed verotoxin 2 gene (vt2) that was used to generate vt2 probe and second primer set for nested PCR to attempt more sensitive detection. Most sensitive nested PCR revealed that 45% of tested cattle and 47% of tested zoo animals were VTEC-positive, while least sensitive normal PCR detected VTEC from none of these animals except a mountain goat. Moderately sensitive detection by PCR in combination with hybridization suggested that the VTEC density varied between the VTEC-positive cattle.

• Journal of Life Science
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• v.10 no.4
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• pp.333-339
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• 2000

To identify genes involved in the decolorization of crystal violet, we isolated random mutants generated by transponson insertion in crystal violet-declorizing bacterium, Citrobacter sp. The resulting mutant bank yielded mutants with six distinct phenotypes, and Southern hybridization with a Tn5 fragment as a probe showed a single hybridized with six distinct phenotypes, and Southern hybridization with a Tn5 fragment as a probe showed a single hybridized band in the mutants Ctg 2, 5 an 6, whereas two and three bands were detected in Ctg1, 4 and 3, respectively. Tn5-inserted genes were isolated and the DNA sequence flanking Tn5 was determined. From comparison with a sequence database, putative protein product encoded by ctg 5 was identified as E. coli maltose transproter(Mal G) homolog, whereas the deduced amino acid sequence of the other ctg genes did not show any significant similarity with any DNA or protein sequency. Therefore, these results indicate that the other ctg genes except ctg 5 encode new proteins responsible for decolorization of crystal violet.

• Journal of Microbiology and Biotechnology
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• v.11 no.1
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• pp.1-6
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• 2001

Ten strains of plasmid-harboring Bifidobacterium sp. were isolated from the feces of adults and children, and named as Bifidobacterium sp. GE1-GE8, ST, and SH5. These plasmids were categorized into three homologous groups (pKJ50-homologous, pKJ36-homologous, and non-homologous groups) according to Southern hybridization patterns using the formerly characterized bifidobacterial plasmids, pKJ50 and pKJ36, as probes. nine strains harboring the plasmids were shown to accumulate single-stranded DNA as a replication intermediate, based on the S1 nuclease treatment and Southern hybridization. These results suggest that the strains replicate by a rolling circle mechanism. Minimal inhibitory concentrations (MIC) of the isolated bifidobacteria against several antibiotics were determined. Two strains, GE2 and GE3, showed relatively high MiC values against tetracycline ($793.6{\mu}g/ml$) and erythromycin ($153.6{\mu}g/ml$), respectively. The tetracycline resistance of GE2 disappeared when the resident plasmid of GE2 was cured by ethidium bromide. These results show that pKJ36-homologous and pKJ50-homologous plasmids are prevalent among various Bifidobacterium strains and some Bifidobacterium plasmids appear to code for antibiotic resistance.

• International Journal of Oral Biology
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• v.38 no.2
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• pp.43-49
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• 2013

The objective of this study was to develop PCR primers that are specific for Streptococcus sanguinis, Streptococcus parasanguinis, and Streptococcus gordonii. We designed the S. sanguinis-, S. parasanguinis-, and S. gordonii-specific primers, Ssa21-F3/Ssa21-R2, Spa17-F/Spa17-R, and Sgo41-F1/Sgo41-R1 respectively, based on the nucleotide sequences of the Ssa21, Spa17, and Sgo41 DNA probes that were screened using inverted dot blot hybridization (IDBH). The species-specificity of these primers was assessed against 43 strains of mitis group streptococci, including clinical strains of S. sanguinis, S. parasanguinis, and S. gordonii. The resulting PCR data revealed that species-specific amplicons had been obtained from all strains of the target species tested, and that none of these amplicons occurred in any other strains from other species. These results suggest that the Ssa21-F3/Ssa21-R2, Spa17-F/Spa17-R, and Sgo41-F1/Sgo41-R1 primers may be useful in detecting S. sanguinis, S. parasanguinis, and S. gordonii at the species level, respectively.

• Korean Journal of Breeding Science
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• v.43 no.4
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• pp.252-259
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• 2011

Conventional staining and fluorescence in situ hybridization (FISH) karyotypes of the non-genetically modified (GM) parental rice line, 'Nakdong' (Oryza sativa L. japonica), and its four GM rice lines, LS28 (event LS30-32-20-1), Cry1Ac1 (event C7-1-9-1), and LS28 ${\times}$ Cry1Ac1 (events L/C1-1-3-1 and L/C1-3-1-1) were analyzed using 5S and 45S rDNAs as probes. Both parental and transgenic lines were diploids (2n=24) with one satellite chromosome pair. The lengths of the prometaphase chromosomes ranged from 1.50 to $6.30{\mu}m$. Four submetacentric and eight metacentric pairs comprised the karyotype of 'Nakdong' and its four GM lines. One pair of 5S rDNA signals was detected near the centromeric region of chromosome g in both the parental and transgenic lines. The 45S rDNA signals were detected on the secondary constrictions of the satellite chromosome pair in both the parental and transgenic lines. There was no significant difference in chromosome size, length, and composition between 'Nakdong' and its four GM lines. This research was conducted as a preliminary study for chromosomal detection of transgenes in GM rice lines and would be useful for their breeding programs.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.5
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• pp.657-662
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• 2008

To develop transgenic orchardgrass (Dactylis glomerata L.) resistant to high temperature, the recombinant DgHSP17.2 gene was introduced into orchardgrass plants using the Agrobacterium-mediated transformation method and expressed constitutively under the control of the CaMV 35S promoter. The results of genomic DNA PCR and Southern analysis showed a DNA band and hybridization signal on agarose gel and X-ray film in transgenic orchardgrass plants harboring the recombinant DgHSP17.2 gene, but a DNA band and hybridization signal were not observed in the wild type and empty vector control plants. The same result was also obtained in RT-PCR and Southern blot analysis, and these transgenic orchardgrass plants did not show any morphological aberration both in the culture bottle and soil mixture. When leaf discs cut from transgenic orchardgrass plants with recombinant DgHsp17.2 gene were exposed to lethal temperature (heat treatment at $60^{\circ}C$ for 50 min), 60-80% of the leaf discs showed only damage symptoms, but non-transgenic leaf discs showed a lethal condition. These results indicate that the DgHsp17.2 gene may act as a protector from heat stress in plants.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.4
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• pp.405-415
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• 2000

Objective: The present study was undertaken to synthesize a human Y chromosome specific probe and to confirm the usefulness of the probe for fluorescence in situ hybridization (FISH) in various types of human cells. Methods: An approximately 400 bp DNA fragment of the DYZ1 sequences was synthesized by PCR using digoxigenin labeled dUTP (dig-PCR). The fidelity of probe was tested by FISH for cultured and uncultured human lymphocytes, amniocytes, chorionic villus cells, embryos, sperms, and germ cells of seminiferous tubule. Results: The human Y chromosome specific probe hybridized specifically to Y chromosome of the cells that had been tested. This probe assigned to the Yq12 region where the DYZ1 repetitive sequence is concentrated. Conclusion: We have identified a human Y chromosome specific probe that hybridized specifically to the Y chromosome by FISH for various types of uncultured as well as cultured cells. Therefore FISH technique using human Y chromosome specific probe should be useful for clinical application as a diagnostic tool for the detection of human Y chromosome.