• Asian-Australasian Journal of Animal Sciences
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• 제23권1호
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• pp.17-24
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• 2010

In the antral follicle phase, several layers of cumulus cells surround the oocyte and play an important support and regulation role in oocyte development and maturation via intercellular communications and interactions between oocytes and cumulus cells. However, information on stage specific gene expression in swine during the phase is not well understood. To investigate the function of cumulus cells during in vitro maturation of porcine oocytes and gene expression, suppression subtractive hybridization (SSH) was performed to screen genes that were differentially expressed between cumulus-oocyte complexes (COCs) and naked oocytes (NOs). Utilizing mRNAs from in vitro maturation oocytes, a SSH cDNA library from COCs as the tester and NOs as the driver was constructed. The SSH cDNA library was then screened using dot blot analysis. Results showed that a total of 70 clones randomly selected from the library were differentially expressed. Among these, 41 exhibited high homology to known genes and 11 were novel expressed sequences tags (ESTs). Four differentially expressed genes, including bfgf, sprouty 2, egr and btc, were further studied by real time quantitative PCR; results confirmed an increased expression of respective mRNA in COCs compared with NOs, which suggests that these factors may play an important role in oocyte development and maturation.

• KSBB Journal
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• 제19권4호
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• pp.308-314
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• 2004

모넨신, 니저리신, 사이클로스포린 등을 하이드록실레이션 시키는 균주인 S. benihana에 존재하는 여러 가지 CYP를 클로닝하기 위해, 방선균 CYP의 보존된 부분을 통해서 degenerate primer를 제작하였고, colony hybridization을 통해서 스크리닝 한 결과 총 5 종류의 CYP가 검색되었다. 아미노산 서열의 분석 결과 방선균의 CYP 들과 매우 높은 유사성을 가졌으며, 이들 CYP의 앞 뒤 서열의 검색 결과 이 중 4개의 CYP의 downstream에는 FD 유전자가 존재함을 알 수 있었다. CYP503의 경우 다른 나머지 4개의 CYP의 서열과 차이가 많았으며, 2차 대사산물의 변형과 관련되어 있을 것으로 예상되며, ChoP와 유사성을 보이는 나머지 4개의 CYP는 스테로이드 계열 물질의 하이드록실레이션과 밀접한 연관이 있을 것으로 추정된다.

• 한국어병학회지
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• 제10권1호
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• pp.39-44
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• 1997

Metallothionein은 세균에서 척추동물에 이르기 까지 모든 생명체에 존재하며, 중금속의 세포내농도를 조절하는 중요한 단백질이다. 현재까지 metallothionein의 기능 및 유도기작에 관한 연구는 많이 진척되지는 않았으나, 여러 metallothionein 유전자의 구조가 밝혀져 있는 실정이다. 특히 어류의 metallothionein은 여러종류의 중금속과 환경적인 자극에 의하여 유도되고 정량적인 RT-PCR의 방법으로 metallothionein 유전자의 RNA transcript를 측정함으로써 환경적인 자극의 정도와 중금 속의 상대적인 양을 측정할 수 있기 때문에 중요한 단백질로 인식되고 있다. 본 연구에서는 유전자내부의 특이적 primer와 통상적인 3`말단의 primer를 이용하여 PCR에 의해 450 bp에 해당하는 metallothionein 유전자의 일부의 특성을 조사하였다. 챠넬메기의 cDNA library로부터 PCR에 의해 증폭된 450 bp의 PCR 절편은 다른 어류의 metallothionein 유전자와는 유사성을 보이지 않았다.

• 한국어병학회지
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• 제10권2호
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• pp.87-95
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• 1997

새우의 갑각에 흰점을 유발하는 특징을 가진 WSBV는 Baculovirus의 일종으로 여러 종류의 새우에 높은 치사율을 보이는 병원체로서 새우양식에 막대한 피해를 주고 있다. 본 연구에서는 국내에서 양식중인 대하에 질병을 유발한 WSBV의 특성을 알아내고자 치사한 새우로부터 바이러스의 게놈을 클로닝하여 재조합클론(E3)을 분자생물학적으로 분석하였다. E3의 염기서열을 분석한 결과, 이 클론은 AcNPV를 포함한 지금까지 알려진 어떠한 바이러스와도 뚜렷한 상동성(60%)을 보이지 않아 WSBV가 기존의 바이러스와 구분되는 새로운 바이러스임을 알 수 있다. E3의 염기서열에 기초하여 한쌍의 PCR 프라이머를 작성하였다. 병든 새우로부터 분리한 DNA를 30회 증폭한 결과, 예상크기의 산물을 얻을 수 있어 이 방법은 바이러스의 감염여부를 알아낼 수 있는 진단법으로도 활용가능하다.

• Journal of Microbiology and Biotechnology
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• 제12권4호
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• pp.544-552
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• 2002

Several microorganisms from rat and human feces and lumen fluid of cows were screened for their ability to decolorize the synthetic dyes. Consequently, a novel dye-degrading strain AB&J was isolated. Taxonomic identification including 165 rDNA sequencing and phylogenetic analysis indicated that the isolate had 99.9% homology in its 165 rDNA base sequence with Clostridium perfringens. After 27 h Incubation with the strain, brilliant blue R, bromophenol blue, crystal violet, malachite green, methyl green, and methyl orange were decolorized by about 69.3%, 97.7%, 96.3%, 97.9%, 75.1%, and 97.2%, respectively. The triphenlmethane dye, bromophenol blue, was decolorized extensively by growing Clostridium perfringens AB&J cells in liquid cultures under anaerobic condition, although their growth was strongly inhibited in the initial stage of incubation. This group of dyes is toxic, depending on the concentration used. The dye was significantly decolorized at a relatively lower concentration of below 50 $\mu g \;ml^{-1}$, however, the growth of the cells was mostly suppressed at a dye concentration of 100 $\mu g \;ml^{-1}$. The decolorization activity in cell-free extracts was much higher in cytoplasm than in periplasm and cytoplasmic membrane. Therefore, the enzyme related uptake of bromophenol blue seemed to be localized in cytoplasm. The optimal pH and temperature of bromophenol blue uptake fur decolorization activities were 7.0 and 4$0^{\circ}C$, respectively.

• Journal of Plant Biotechnology
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• 제34권3호
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• pp.173-180
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• 2007

The full-length cDNA of LeAPR1 encoded a protein of 461 amino acid residues, which contained homology with phosphoadenosine phosphosulphate reductase (PAPS reductase) in N-terminal and an adenylylsulfate reductase in N-term and C-terminal. Analysis of the deduced amino acid sequence of LeAPR1 revealed that it shares high sequence identity with potato StAPR (96% identity)(Gene bank accession no. CDC44841). We found that multiple copies of LeAPR1 gene are present in the tomato genome through southern blot using genomic DNA was digested with 3 different restriction enzymes. The expression of LeAPR1 was also examined in various organs and its expression was also detected at high levels in roots and stems. Only high amounts of LeAPR1 transcripts were detected at high transcripts in the leaves at time 0, and then reduced as the plant stressed by the NaCl and abscisic acid (ABA). After 24h treatment of NaCl and ABA were showed increasing patterns of LeAPR1 gene. Time course of LeAPR1 gene expression was examined under oxidative stresses from metyl viologen (MV) and hydrogen peroxide ($H_2O_2$). In the presence of 10 mM $H_2O_2$ and $50\;{\mu}M$ MV, the levels of LeAPR1 transcript in leaves decreased after 1 h, and then increased strongly, peaked at 24 h. Our results indicated that LeAPR1 may play a role function of circadian regulation involved in abiotic stresses signaling pathways.

• The Plant Pathology Journal
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• 제26권1호
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• pp.8-16
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• 2010

The phytopathogenic fungus Magnaporthe oryzae is a major limiting factor in rice production. To understand the genetic basis of M. oryzae pathogenic development, we previously analyzed a library of T-DNA insertional mutants of M. oryzae, and identified ATMT0879A1 as one of the pathogenicity-defective mutants. Molecular analyses and database searches revealed that a single TDNA insertion in ATMT0879A1 resulted in functional interference with an annotated gene, MGG00056, which encodes a short-chain dehydrogenase/reductase (SDR). The mutant and annotated gene were designated as $MoSDR1^{T-DNA}$ and MoSDR1, respectively. Like other SDR family members, MoSDR1 possesses both a cofactor-binding motif and a catalytic site. The expression pattern of MoSDR1 suggests that the gene is associated with pathogenicity and plays an important role in M. oryzae development. To understand the roles of MoSDR1, the deletion mutant ${\Delta}Mosdr1$ for the gene was obtained via homology-dependent gene replacement. As expected, ${\Delta}Mosdr1$ was nonpathogenic; moreover, the mutant displayed pleiotropic defects in conidiation, conidial germination, appressorium formation, penetration, and growth inside host tissues. These results suggest that MoSDR1 functions as a key metabolic enzyme in the regulation of development and pathogenicity in M. oryzae.

• International Journal of Industrial Entomology and Biomaterials
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• 제27권1호
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• pp.159-165
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• 2013

Bone morphogenetic proteins (BMPs) belong to the transforming growth factor (TGF-${\beta}$) superfamily and are involved in osteoblastic differentiation. The largest TGF-${\beta}$ superfamily subgroup shares genetic homology with human BMPs (hBMPs) and silkworm decapentaplegic (dpp). In addition, hBMPs are functionally interchangeable with Drosophila dpp. Bombyx mori dpp may induce bone formation in mammalian cells. To test this hypothesis, we synthesized the 1,285-base pairs cDNA of full-length B. mori dpp using total RNAs obtained from the fat body of 3-day-old of the $5^{th}$ instar larvae and cloned the cDNA into the pCEP4 mammalian expression vector. Next, B. mori dpp was expressed in C3H10T1/2 cells. The target cells transfected with the pCEP4-Bm dpp plasmid showed biological functions similar to those of osteogenic differentiation induction growth factors such as hBMPs. We determined the relative mRNA expression rates of Runt-related transcription factor 2 (RUNX2), osterix, osteocalcin, and alkaline phosphatase (ALP) to validate the osteoblast-specific differentiation effects of B. mori dpp by performing quantitative real-time RT-PCR. Interestingly, mRNA expression levels of the 3 marker genes except RUNX2, in cells expressing B. mori dpp were much higher than those in control cells and C3H10T1/2 cells transfected with pCEP4. These results suggested that B. mori dpp signaling regulates osterix expression during osteogenic differentiation via RUNX2-independent mechanisms.

• Asian Journal of Atmospheric Environment
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• 제5권3호
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• pp.164-171
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• 2011

This work focuses on the analysis of bioaerosols in the atmosphere at higher altitudes over Noto Peninsula, Japan. We carried out direct sampling via aircraft, separated cultures, and identified present isolates. Atmospheric bioaerosols at higher altitudes were collected using a Cessna 404 aircraft for an hour at an altitude of 3,500 m over the Noto Peninsula. The aircraft-based direct sampling system was devised to improve upon the system of balloon-based sampling. In order to examine pre-existing microorganism contamination on the surface of the aircraft body, bioaerosol sampling was carried out just before takeoff using the same method as atmospheric sampling. Identification was carried out by a homology search for 16S or 18S rDNA isolate sequences in DNA databases (GenBank). Isolate sampling just before takeoff revealed Stretpomyces sp., Micrococcus sp., and Cladosporium sp. One additional strain, Bacillus sp., was isolated from the sample after bioaerosol collection at high altitude. As the microorganism contamination on the aircraft body before takeoff differed from that while in the air, the presence of additional, higher atmosphere-based microorganisms was confirmed. It was found that Bacillus sp. was floating at an altitude of 3,500 m over Noto Peninsula.

• Journal of Microbiology and Biotechnology
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• 제12권3호
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• pp.437-443
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• 2002

A number of soil and wastewater samples were collected from the vicinity of an effluent treatment plant for the chemical industry. Several microorganisms were screened fur their ability to decolorize the triphenylmethane group of dyes. As a result, a novel crystal violet dye-degrading strain LK-24 was isolated. Taxonomic identification including 16S rDNA sequencing and phylogenetic analysis indicated that the isolate had a $99.5\%$ homology in its 16S rDNA base sequence with Stenotrophomonas maltophilia. The triphenylmethane dye, crystal violet, was degraded extensively by growing cells of Stenotrophomonas maltophilia LK-24 in agitated liquid cultures, although their growth was strongly inhibited in the initial stage of incubation. This group of dyes is toxic, depending on the concentration used. The dye was significantly degraded at a relatively lower concentration, below $100{\mu}g\;ml^-1$, yet the growth of the cells was totally suppressed at a dye concentration of $250{\mu}g\;ml^-1$. The degradation products of crystal violet were identified as 4,4'-bis(dimethylamino)-benzophenone and ${\rho}$-dimethylaminophenol by Gas chromatography-Mass spectrometry. The 4,4'-bis(dimethylamino)-benzophenone was easily obtained in a reasonable yield, as it was not metabolized further by S. maltophilia LK-24; however, the ${\rho}$-dimethylaminophenol was not easily identifiable, as it was further metabolized.