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Rapid, sensitive and specific detection of Brucella canis by real-time PCR assay using HybProbe
( Sung Ll Kang ),( Moon Her ),( Ji Yeon Kim ),( Kichan Lee ),( Jong Wan Kim ),( Hyang Keun Lee ),( So Ra Sung ),( Hyorim Cho ),( Suk Chan Jung ) 한국예방수의학회(구 한국수의공중보건학회) 2013 예방수의학회지 Vol.37 No.1
We designed and evaluated a specific HybProbe set for identification of Brucella canis, which is based on a specific single nucleotide polymorphism (SNP) of the omp25 gene. Real-time PCR assay using this probe specifically identified B. canis reference and field strains, however, the other Brucella species and general bacteria strains were not amplified. This real-time PCR assay can re-confirm specific amplification in melting curve analysis. The average melting temperature values for generation of B. canis strains wan 67℃m, due to a single base-pair mismatch at the binding site of the probe. The detection limit appeared to be 30 fg of DNA and 100CFU by serial dilution of whole blood inoculated with B. canis. In addition, use of the buffy coat resulted in approximately 50 times higher sensitivity than whole blood directly. This technique is widely applicable for use in diagnostic laboratories and is readily discriminative with regard to small amounts of biomass.
Lee, Chae-Kwan,Kang, Han-Seung,June, Bu-ll,Lee, Byung-Ju,Moon, Deog-Hwan,Kang, Sung-Goo The Korean Society for Integrative Biology 2001 Korean journal of biological sciences Vol.5 No.4
There is a critical developmental period during which brain sexual differentiation proceeds irreversibly under the influence of gonadal hormone. Recently, kinesin superfamily-associated protein 3 (KAP3) gene expressed during the 'critical period' of rat brain differentiation was identified by us (Choi and Lee, 1999). KAP3 functions as a microtubule-based motor that transports membranous organelles anterogradely in cells, including neurons (Yamazaki et al., 1996). mRNA level of KAP3 gene markedly increased before the initiation of puberty. Neonatal treatment of estrogen clearly inhibited the prepubertal increase in KAP3 mRNA level (Choi and Lee, 1999). In the present study, we aimed to investigate the effects of polychlorinated biphenyls (PCBs), as endocrine disruptors (EDs) on the expression of KAP3 gene during the 'critical period' of rat brain development. In our data, PCBs significantly decreased the expression of KAP3 gene in the fetal (day 17) and the neonatal (day 6 after birth in) male and female rat brains. The body weight and the breeding ability were significantly decreased in the PCBs-exposed rats compared with the control. These results showed that PCBs affect the transcriptional level of brain sexual differentiation related gene, KAP3, in the fetal and the neonatal rat brains. The maternal exposure to the PCBs may lead to toxic response in embryonic brain sexual differentiation and breeding ability after sexual maturation. This study indicates that KAP3 gene may be useful as a gene marker to analyze the molecular mechanism of toxic response in the animal brain development and sexual maturation exposed to PCBs.
Immunological analyses for recombinant GroEL of Brucella canis
( Hyang Keun Lee ),( Jong Wan Kim ),( Kichan Lee ),( Moon Her ),( Ji Yeon Kim ),( Sung Ll Kang ),( Suk Chan Jung ) 한국예방수의학회(구 한국수의공중보건학회) 2013 예방수의학회지 Vol.37 No.1
Brucella(B.) canis is transmitted primarily by direct contact with aborted fetuses and placenta. It is also known to infect humans, which likely results in infectious risk to veterinarians and companion animal owners. For development of an effective subunit vaccine against canine brucellosis, immunogenicity for chaperonin GroEL(heat shock protein of 60kDa) of B. canis was investigated by recombinant protein vaccine, which is known to induce humoral and cellular immune responses in mouse models. Mice received subcutaneous injection in the lower back midline with 30 ug of purified GroEL on days 0, 14. Sera were obtained at 0, three and six weeks after the first immunization for detection of humoral and cellular immunity. Significantly higher lgG1 and lgG2a were detected in the sera of mice immunized with recombinant GroEL, compared to those inoculated with negative control.