• 미생물학회지
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• 제42권1호
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• pp.12-18
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• 2006

2003년부터 2004년까지 정북지역에서 발생한 설사환자로부터 17주, 돼지로부터 18주 등 총 35주의 Salmonella enterica serovar Typhimurium이 분리되었고, 분리된 균주를 대상으로 항균제 내성양상, class I integron 특성 및 pulse-field gel electrophoresis유형(PFGE 유형; pulsotype)이 조사되었다. 35주 모두가 한가지 이상의 항균제에 내성을 나타내었고, 돼지로부터 분리된 대부분의 균주는 ampicillin, chloramphenicol, streptomycin, sulfamethoxazole/trimethoprime, tetracyclin, nalidixic acid에 내성을 나타내었다. 35주를 대상으로 class I, II 및 III integron gene cassette를 검색한 결과, 설사환자로부터 분리된 17주 중 3주가 dhfrX-orfF-aadA2 integron gene cassette을 보유하였고, 돼지로부터 분리된 18주중 11주가 dhfrX-orfF-aadA2 integron gene cassette를, 1주가 aadA2 integron을 보유하였다. 그러나 35주 모두가 class 2 integron gene cassette와 class 3 integron gene cassette는 보유하지 않은 것으로 나타났다. 35주의 PEGE 유형은 5가지로 분류되었으며, 31주가 pulsotype A로 나머지 4주는 pulsotype B, C, D, E형으로 각각 나뉘어졌다. 이러한 결과로 볼 때 경북지역에서 사람과 돼지에서의 S. enterica serovar Typhimurium에 의한 살모넬라중은 몇 종류 유행주의 전파에 의한 것임을 보여줄다. dhfrX-orfF-aadA2 integron gene cassette를 보유한 13주가 pulsotype A, dhfrX-orfF-aadA2 integron gene cassette을 보유한 1주가 pulsotype B, aadA2 integron을 보유한 1주가 pulsotype E, integron을 보유하고 있지 않은 15주가 pulsotype A로 나타났다.

• 한국임상수의학회지
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• 제21권2호
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• pp.102-108
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• 2004

Orf virus (ORFV), a member of genus Parapoxvirus (family-Poxviridae), a causative agent of contagious ecthyma in sheep and goat leading to a condition commonly known as vesicular dermatitis. Recently, twelve goats from Iksan in Jeonbuk province were observed with clinical signs like necrotic vesicular lesions around the mucosa of mouth, nasal cavity, eye, ear, teats, abdomen and groin. Based on these clinical symptoms, contagious ecthyma infection was suspected. The skin scrapping was collected from lesions for isolation of DNA and subsequent PCR amplification of ORFV specific 235 bp region of B2L gene. All of the samples were found positive by PCR analysis. Sequencing and further phylogenetic analysis of the PCR product revealed 100% identity to Japan isolate of ORFV (Okinawa, GenBank accession number AB080769), and showed 99.6% of similarity to New Zealand strain (NZ-2, GenBank accession number U06671). It was concluded that ORFV strain detected in the present study is homologous to Japan isolate and New Zealand strain. The PCR test based on amplification of B2L gene is a highly useful tools for rapid and specific diagnosis of contagious ecthyma.

• Journal of Microbiology
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• 제37권2호
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• pp.86-89
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• 1999

An OTHBVS cell line from HepG2 was established. This cell line stably expresses the human hepatitis B virus (HBV) middle S protein that includes the preS2 region which is important for HBV particle entry into the hepatocyte. To establish this cell line, the middle S open reading frame (ORF), with a promoter located in the 5' region and enhancer located in the 3' region, was cloned downstream from the metallothionine (MT) promoter of the OT1529 vector. In this vector, expression of the middle S protein was constructed to be regulated by its own promoter and enhancer. Expression of the large S protein which contains the preS1 region in addition to the middle S protein was designed to be regulated by the MT promoter. When extracts of OTHBVS cells were examined with an S protein detection kit (RPHA, Korea Green Cross Co.), an S protein was detected. Total mRNA of OTHBVS cell examined by northern blot analysis with an S ORF probe revealed small/middle S transcripts (2.1 kb). When the MT promoter was induced by Zn, large S transcripts (2.4 kb) were detected. The GP36 and GP33 middle S proteins were presumably detected, but large S proteins were not detected by immunostain analysis using anti-preS2 antibody.

• 원예과학기술지
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• 제32권2호
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• pp.261-268
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• 2014

Senescence of Hosta ventricosa flowers was firstly characterized as ethylene-sensitive since the deterioration of the tepal was accompanied by increased endogenous ethylene biosynthesis. The full-length cDNAs and DNAs of 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS) and ACC oxidase (ACO) involved in ethylene biosynthesis were cloned from H. ventricosa flowers. The HvACS ORF with 1347 bp and two introns, encoded a polypeptide of 448 amino acids showing 79% homology with that in Musa acuminata. The HvACO ORF contained 957 bp and three introns, encoding a 318-residue polypeptide showing 83% homology with that in Narcissus tazetta. The timing of the induction of HvACS expression was in correspond to the timing of the increase in ethylene production, and that the up-regulation of HvACO transcript was closely correlated with an elevated ethylene production, but underwent a down-regulation in wounded leaves with elevated ethylene emission. The results, together with expression analysis in vegetative tissues, suggested that both HvACS and HvACO were specifically regulated by flower senescence.

• 미생물학회지
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• 제30권6호
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• pp.456-459
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• 1992

Halobacteria 의 gvpD. gvpE 유전자는 가스포 형성에 관여하는 유전자로, 이들은 그 transcripts 의 분석에 있어 특유의 연약성 때문에 많은 실험상의 문제를 야기시키고 있다. 본 실험은 연약한 mRNA 를 reverse transcriptase 를 사용, DNA 로 바꾸고 이를 다시 PCR(Polymerase Chain Reaction) 방법으로 증폭시킴으로써, 유전자의 연약한 mRNA 를 다시 상보적인 안정한 DNA 로 대치케 하여 RNA 상의 cloning, RNA sequencing 을 용이하게 하였다. 결과는 유전자 gvpD 에서 거의 전 ORF(Open Reading Frame) 의 범위에서 northern hybridization 에서 발견치 못한 transcipts 를 확인할 수 있었다.

• BMB Reports
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• 제39권6호
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• pp.737-742
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• 2006

Open reading frame 60 of Bombyx mori nucleopolyhedrovirus (Bm60) is located between 56,673 and 57,479 bp in the BmNPV genome which encodes 268 amino acid residues with predicted molecular weight of 31.0 kDa. Bm60 and its homologues have been identified in 11 completely sequenced lepidopteran NPVs. The transcript of Bm60 was detected by RT-PCR at 18-72 h post-infection (p.i.), while the corresponding protein could be detected at 24-72 h p.i. in BmNPV-infected BmN cells by Western blot analysis using a polyclonal antibody against Bm60. The expression of Bm60 was inhibited in the presence of Ara-c, an inhibitor of viral DNA synthesis. These results together indicated that Bm60 was a late gene. The size of Bm60 product was found to be a 31 kDa in BmNPV-infected BmN cells, consistent with predicted molecular weight. Immuno-fluoresence analysis showed that the Bm60 product was first detected in the cytoplasm at 24 h p.i and also located in nucleus during later infection. In conclusion, the available data suggest that Bm60 is a functional ORF of BmNPV and encodes a 31kDa protein expressed in the later stage of infection cycle.

• Asian Pacific Journal of Cancer Prevention
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• 제15권19호
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• pp.8051-8055
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• 2014

Background: A common genetic variant rs3757318, located in intron of C6orf97, was firstly identified to be associated with breast cancer (BC) risk by a genome-wide association (GWA) study. However, subsequent validation studies with different ethnicities have yielded conflicting results. Materials and Methods: We performed a meta-analysis to synthesize all available data for evaluating the precise effect of this variant on BC susceptibility. Results: A total of 8 articles containing 11 studies with 62,891 cases and 65,635 controls were included in this meta-analysis. When compared to the G allele, the rs3757318-A allele was significantly associated with BC risk with the pooled OR of 1.21 (95% CI=1.15 - 1.29, P<0.001) but with obvious between-study heterogeneity (P=0.040). Stratified analysis suggested that diversity of ethnicity along with control source may explain part of the heterogeneity. Similarly, significant associations were also identified in heterozygote, homozygote, dominant and recessive genetic models. Sensitivity and publication bias analyses indicated robust stability of our results. Conclusions: Our present meta-analysis demonstrated that the variant rs3757318 is associated with increased BC risk. Nevertheless, further studies are needed to clarify the underlying biological mechanisms.

• BMB Reports
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• 제43권5호
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• pp.337-343
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• 2010

Fenofibrate, an agonist of $PPAR{\alpha}$, plays an important role in activating many proteins catalyzing lipid metabolism, and it also has a considerable effect on improvement of insulin sensitivity in the diabetic condition. To investigate fenofibrate-dependent expression of peripheral tissue proteins in diabetes, we analyzed whole muscle or fat proteins of fenofibrate-fed OLETF rats, an animal model of type II diabetes, using 2-dimensional gel electrophoresis. We found that many proteins were specifically expressed in a fenofibrate-dependent manner in these diabetic rats. In particular, a functionally unknown C11orf59 protein was differentially expressed in the muscle tissues (about 5-fold increase) in fenofibrate-fed OLETF rats as compared to control rats. Additionally, the signal proteins phosphatidylethanolamine binding protein and IkB interacting protein were differentially regulated in the fenofibrate-treated adipose tissues. We suggest here that these proteins might be involved in controlling lipid or carbohydrate metabolism in diabetes via $PPAR{\alpha}$ activation.

• Journal of Microbiology and Biotechnology
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• 제16권3호
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• pp.450-456
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• 2006

Pseudomonas fluorescens MC07 is a growth-promoting rhizobacterium that suppresses mycelial growth in fungi such as Rhizoctonia solani, Pythium ultimum, Fusarium oxysporum, and Phytophthora capsici. To determine the role of the bacterium's antifungal activity in disease suppression, we screened 2,500 colonies generated by Tn5lacZ insertions, and isolated a mutant 157 that had lost antifungal activity. The EcoRI fragment carrying Tn5lacZ was cloned into pBluescript II SK(+) and used as a probe to isolate wild-type clones from a genomic library of the parent strain, MC07. Two overlapping cosmid clones, pEH4 and pEH5, that had hybridized with the mutant clone were isolated. pEH4 conferred antifungal activity to the heterologous host P.fluorescens strain 1855.344, whereas pEH5 did not. Through transposon mutagenesis of pEH4 and complementation analyses, we delineated the 14.7-kb DNA region that is responsible for the biosynthesis of an antifungal compound. DNA sequence analysis of the region identified 11 possible open reading frames (ORF), ORF1 through ORF11. A BLAST search of each putative protein implied that the proteins may be involved in an antifungal activity similar to polyketides.

• 대한수의학회지
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• 제38권2호
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• pp.304-313
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• 1998

Sixteen Korean field transmissible gastroenteritis viruses (TGEVs) were isolated using swine testicular cell (STC) and the genomic diversity of them was analyzed. All TGEV isolates produced a typical cytopathic effect in STC and were confirmed as TGEV by immunofluorescence assay using monoclonal antibody against TGEV and PCR using TGEV specific primers. RNAs from TGEV field isolates and vaccine TGEV were extracted and amplified by RT and PCR. The RT-PCR products were digested with selected restriction enzymes and analyzed RFLP patterns. The N-terminal end region of S gene and ORF 3 and 3-1 genes of TGEV amplified by TGEV specific primer pairs seemed to be conserved. Most specific variations were detected in S gene amplified by TGEV 4/6 primer pairs which includes antigenic sites A and D. When the PCR products were treated with Sau3AI and Ssp I, Bvac(vaccine strain), field isolates 133 and 347 were differentiated from Miller and Purdue types. In the case of D5 field isolates, it was classified into Purdue type by Sau 3AI but classified into independent TGEV by Ssp I. Two different TGEV strains from D2 sample were confirmed by plaque purification and RT-PCR-RFLP analysis. To investigate the change occurring in TGEV genome after serial passage, the TGEV P44 strain was passaged through STC. There were specific changes in S gene and a large deletion was observed in ORF 3 and 3-1 genes. These studies showed that a distinct difference in genome exists among TGEV field isolates.