• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.602-605
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• 2001

Low-temperature adaptation and protection for environmental stresses were studied in the gram-negative soil bacterium Bradyrhizobium japonicum 61A101c. B. japonicum was more resistant to alcohol, $H_2O_2$, heat and freezing following a pretreatment at $4^{\circ}C$, resulting in approximately 10 to 1,000 folds increased survival compared to mid-exponential-phase cells grown at an optimal temperature at $28^{\circ}C$. This phenomena relate to the cold shock protein expressed when cells are exposed to a downshift in temperature. To confirm the presence of cold shock protein genes in B. japonicum, a PCR strategy was employed using a degenerate primer set, which successfully amplified a putative csp gene fragment. Sequence analysis of the PCR product(200bp) revealed csp-like sequences that were up to 96% identical to csp gene of S. typhimurium.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.827-828
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• 2001

We developed a geno-chromatographic assay system based on hybridization between target DNA (e.g., amplified product from PCR) and a capture probe immobillized on a predetermined site of membrane. This system can offer a rapid detection of a gene sequence specific to a certain type of cells as well as simplicity of the analytical procedure. Such performances were demonstrated by utilizing Salmonella typhimurium as model analyte.

• BMB Reports
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• v.30 no.1
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• pp.33-36
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• 1997

Age-dependent deletion of mitochondrial DNA (mtDNA) was detected in mouse brain using PCR method. The size of the deleted fragment was 0.5 kb, 0.9 kb. 1.7 kb and 4.3 kb in the region between cytochrome b gene and ATPase 6 gene. The deleted fragment was increased gradually from 3-month to 22month Direct repeat sequence flanking the deletion in 0.5 kb PCR product was TAAT.

• Journal of Microbiology and Biotechnology
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• v.16 no.8
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• pp.1185-1191
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• 2006

The expression of a pullulanase gene in Pichia pastoris was investigated. The gene encoding pullulanase was cloned by PCR using the chromosomal DNA of Bacillus naganoensis as the template. The expression vector pPIC9K-Pu was constructed by inserting the pullulanase gene into plasmid pPIC9K and then transformed into Pichia pastoris SMD 1168 by electroporation. Activity determination, SDS-PAGE, and PCR amplification indicated that the gene of the pullulanase from B. naganoensis had successfully been expressed in SMD 1168 and the molecular size of the expressed recombinant product was about 119.9 kDa. This is the first report on the successful expression of the pullulanase from B. naganoensis in P. pastoris. The transformant secreted recombinant pullulanase with the activity of 350.8 IU/ml in shake-flask culture. The properties of the recombinant pullulanase were characterized.

• Parasites, Hosts and Diseases
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• v.40 no.1
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• pp.25-31
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• 2002

We describe a riboprinting scheme for identification of unknown Acanthamoeba isolates at the species level. It involved the use of PCR-RFLP of small subunit ribosomal RNA gene (riboprint) of 24 reference strains by 4 kinds of restriction enzymes. Seven strains in morphological group I and III were identified at species level with their unique sizes of PCR product and riboprint type by Rsa 1. Unique RFCP of 17 strains in group II by Dde I. Taq I and Hae III were classified into: (1) four taxa that were identifiable at the species level. (2) a subgroup of 4 taxa and a pair of 2 taxi that were identical with each other. and (3) a species complex of 7 taxa assigned to A. castellanii complex that were closely related. These results were consistent with those obtained by 18s rDNA sequence analysis. This approach provides an alternative to the rDNA sequencing for rapid identification of a new clinical isolate or a large number of environmental isolates of Acanthamoeba.

• Journal of Microbiology and Biotechnology
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• v.18 no.12
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• pp.1953-1957
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• 2008

Recombinant Lactobacillus strains have been constructed using gene splicing by overlap extension (SOE). Primers were designed of which one end of an amplified product contained complementary sequences for an end of other amplified fragment. For efficient matching, we used an asymmetric PCR step that was effective at generating an excess of strands that would anneal in the final PCR. CP12, a recombinant fragment consisting of the integrase gene and attachment site of the bacteriophage A2, was constructed and inserted into the genome of Lactobacillus casei ATCC 393, yielding Lb. casei ATCC 393::XCP12. Another recombinant Lb. casei strain was constructed, where the egfp gene was a part of the construction. The EGFP produced from Lb. casei ATCC 393::XCEGFP14 was detected by Western blot hybridization. This simple and widely applicable approach has significant advantages over standard recombinant DNA techniques for Lactobacillus species.

• Journal of Photoscience
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• v.4 no.3
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• pp.141-145
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• 1997

Polymerase chain reaction(PCR) was conducted with a pea cDNA library using two primers synthesized from homology analysis of amino acid sequences for animal and plant cytosolic FBPases. A PCR product with 650 bp long was cloned into pGEM-T vector and sequenced. The deduced amino acid sequence of the cDNA fragment was 98, 91, and 85% homologous with those of cytosolic FBPases from spinach, sugarbeet, and sugarcane, respectively. It was 51% homologous with amino acid sequence of FBPase from pea chloroplasts. Northern blot analysis was proceeded with the cDNA clone resulting that 1.2 kb transcript was highly expressed in light-grown pea leaves but almost not expressed in dark-grown etiolated pea seedlings. When peas grown in the light for 10 days were transferred to darkness, the transcript was gradually decreased with dark treatment, indicating that the expression of the enzyme was induced by continuous white light but suppressed by dark treatment. Pea cytosolic FBPase was highly expressed in leaves with trace amounts in stems. but almost not expressed in roots.

• Journal of Life Science
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• v.10 no.4
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• pp.380-387
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• 2000

Recently Vibrio sp. are the most frequently isolated microorganism, which causing food poisoning. We investigated the detection of toxin genes and effect of chitosan to toxin genes with PCR. Thirty strains of Vibrio sp. were isolated from sea water and sea products through biological and biochemical tests. Out of 30 strains, 8 were identified as V. parahemolyticus, 7 as V. mimcus, 6 as V. damsela, 5 as V. vulnificus, 4 as V. alginoyticus. In detection of ctx, tdh, and t고 as food poisoning-causing toxin genes, ctx from 7 strains, trh from 4 strains and tdh from 6 strains were detected. Among toxin genes detected strains, we selected V. vulnificus-2 V. damsela-1 and V. parahemolyticus-7. As adding chitosan solution to PCR product of 3 strains, the amplified DNA bands were not detected over 450$\mu\textrm{g}$/$m\ell$ concentrantion of chitosan. Over the result, chitosan is thought to influence the detection of toxin gene.

• Korean Journal of Veterinary Research
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• v.39 no.2
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• pp.353-358
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• 1999

New emerging post weaning multisystemic wasting syndrome (PMWS) threatening swine industry worldwide and investigation of the etiological agent is underway. The porcine circovirus (PCV) consistently identified from PMWS pigs and research result indicate that there is strong relationship between PCV and PMWS. Fauns with PMWS submitted pigs suffered from various PMWS typical signs and necropsy finding showed lymph node anomalies. The PCV DNA was amplified from inguinal lymph node collected from pigs with PMWS. PCV specific primers were successfully amplified PCV DNAs and were able to differentiate PCV type I and II. We have identified noble PCV virus with genetic variation. The virus showed insertion of the nucleic acid at the 5' of the genome but did not have PCR product with primer set corresponding to PCV type II virus.

• Korean Journal of Medicinal Crop Science
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• v.17 no.1
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• pp.26-32
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• 2009

The region containing 18S rRNA gene, ITS 1 and part of the 5.8S rRNA gene of the Atractylodes japonica Koidz was amplified by PCR and the product cloned in a pBluescript SK II plasmid. DNA sequence of the cloned DNA was determined and submitted to the GenBank (accession number EU678363). Phylogenetic analysis of the ITS 1 DNA showed close similarity with the other plant species of the family Compositae. The extract of the plant materials of five different members of the family Compositae was analyzed by HPLC to detect atractylon. Extract of the A. japonica Koidz showed presence of significant amount of atractylon. However, noticeable amount of atractylon was not detected by the same analyses from the extracts of the other plants belonging to the family Compositae including Artemisia capillaris, Chrysantemum zawadskii, Eclipta prostrata or Taraxacum platycarpum.