• 식물병연구
• /
• 제18권4호
• /
• pp.387-390
• /
• 2012

2011년에 고창 옥수수 재배지에서 RBSDV에 의한 벼 검은줄오갈병이 발생하였다. 병징은 키가 작아지고 잎에 줄무늬가 형성되면서 심하면 거의 수확이 되지 않는다. 이병주로부터 직접 dsRNA를 추출하여 RT-PCR에 의해 감염을 확인하였다. S8 및 S10 분절의 전 게놈에 특이적인 primer를 이용하여 유전자를 증폭하였다. 이것을 크로닝한 후 전 염기서열을 결정하였다. 염기서열 분석결과 S8 및 S10의 전염기서열은 각각 1,936 nt 및 1,801 nt로 구성되었다. 이는 벼에서 분리된 RBSDV와 같은 크기로 확인되었다. 벼에서 분리된 RBSDV와 상동성 분석결과 S8은 94.9-99.6%, S10은 94.1-98.4%로 나타났다. Phylogenetic tree 분석결과 옥수수에서 분리한 RBSDV S8 및 S10 모두 벼에서 분리된 RBSDV와 같은 그룹으로 분류되었다.

• 생명과학회지
• /
• 제17권8호통권88호
• /
• pp.1129-1134
• /
• 2007

bPDI가 ER내 misfolding 단백질의 생성을 제한함으로써 곤충변역과 관계하는지를 해석하기 위하여 bPDI가 과발현(overexpression)되는 곤충세포주와 이와 반대로 bPDI가 억제발현(knock-down)되는 곤충세포주를 제작하여 bPDI가 곤충면역에 관련하는지를 해석하였다. bPDI가 과발현되는 세포주 (Sf9-bPDI)는 정상세포주(Sf9)나 pIZT/V5-His 벡터만 도입된 세포주(Sf9-pIZT)에 비하여 생존율이 30% 이상 높았지만, bPDI의 전사체 발현이 억제된 세포주(Sf9-bPDI-dsRNA)는 오히려 정상세포주나 pIZT/V5-His 벡터만 도입된 세포주에 비하여 생존율이 약 15%낮았다. 이와 같은 결과로써, bPDI는 ER내 misfolding 단백질의 생성을 제한함으로써 곤충의 ERSE과 밀접하게 관련할 것이라 추정할 수 있었다.

• Animal cells and systems
• /
• 제8권3호
• /
• pp.213-220
• /
• 2004

MicroRNA and siRNA (small interfering RNA), representative members of small RNA, exert their effects on target gene expression through association with protein complexes called miRNP (microRNA associated ribonucleoproteins) and RISC (RNA induced silencing complex), respectively. Although the protein complexes are yet to be fully characterized, human EIF2C2 protein has been identified as a component of both miRNP and RISC. In this report, we raised antiserum against EIF2C2 in order to begin understanding the protein complexes. An immunoblot result indicates that EIF2C2 protein is ubiquitously expressed in a variety of cell lines from human and mouse. EIF2C2 protein exists in both cellular compartments, as indicated by an immunoblot assay with a nuclear extract and a cytosolic fraction (S100 fraction) from HeLa S3 lysate. Depletion of EIF2C1 or EIF2C2 protein resulted in a decrease of microRNA, suggesting a possible role of these proteins in microRNA stability or biogenesis. We also prepared antiserum against dsRNA binding protein PACT, whose homologs in C. elegans and Drosophila are known to have a role in the RNAi (RNA interference) pathway. The expression of PACT protein was also observed in a wide range of cell lines.

• The Plant Pathology Journal
• /
• 제17권3호
• /
• pp.169-173
• /
• 2001

Virus disease occurred up to 62% in average in the greenhouse production of table tomato Seokwang in Suwon, Korea. From symptomatic transition of the labeled tomatoes, two different symptoms, mosaic and bud necrosis, were developed independently. Cucumber mosaic virus necrosis strain (CMV-N) was isolated from table tomato showing bud necrosis symptoms. The isolate caused the bud necrosis on four tomato cultivars and locally infected Chenopodium spp. and Vicia faba by mechanical inculation. The 5th RNA segment, satellite RNA, was identified from CMV-N-infected plants by dsRNA analysis. Crystals of virus particles were observed in cytosols and vacuoles. The virus particles of CMV-N presented abundantly in xylem vessel.

• Parasites, Hosts and Diseases
• /
• 제54권4호
• /
• pp.519-525
• /
• 2016

To investigate the potential role of transforming growth factor (TGF)-${\beta}1$ in liver fibrosis during Echinococcus granulosus infection, 96 BALB/c mice were randomly divided into 2 groups, experimental group infected by intraperitoneal injection with a metacestode suspension and control group given sterile physiological saline. The liver and blood samples were collected at days 2, 8, 30, 90, 180, and 270 post infection (PI), and the expression of TGF-${\beta}1$ mRNA and protein was determined by real-time quantitative RT-PCR and ELISA, respectively. We also evaluated the pathological changes in the liver during the infection using hematoxylin and eosin (H-E) and Masson staining of the liver sections. Pathological analysis of H-E stained infected liver sections revealed liver cell edema, bile duct proliferation, and structural damages of the liver as evidenced by not clearly visible lobular architecture of the infected liver, degeneration of liver cell vacuoles, and infiltration of lymphocytes at late stages of infection. The liver tissue sections from control mice remained normal. Masson staining showed worsening of liver fibrosis at the end stages of the infection. The levels of TGF-${\beta}1$ did not show significant changes at the early stages of infection, but there were significant increases in the levels of TGF-${\beta}1$ at the middle and late stages of infection (P<0.05). RT-PCR results showed that, when compared with the control group, TGF-${\beta}1$ mRNA was low and comparable with that in control mice at the early stages of infection, and that it was significantly increased at day 30 PI and remained at high levels until day 270 PI (P<0.05). The results of this study suggested that increased expression of TGF-${\beta}1$ during E. granulosus infection may play a significant role in liver fibrosis associated with E. granulosus infection.

• The Plant Pathology Journal
• /
• 제34권3호
• /
• pp.191-198
• /
• 2018

Fast and accurate diagnosis is needed to eradicate and manage economically important and invasive diseases like fire blight. Loop-mediated isothermal amplification (LAMP) is known as the best on-site diagnostic, because it is fast, highly specific to a target, and less sensitive to inhibitors in samples. In this study, LAMP assay that gives more consistent results for on-site diagnosis of fire blight than the previous developed LAMP assays was developed. Primers for new LAMP assay (named as DS-LAMP) were designed from a histidine-tRNA ligase gene (EAMY_RS32025) of E. amylovora CFBP1430 genome. The DS-LAMP amplified DNA (positive detection) only from genomic DNA of E. amylovora strains, not from either E. pyrifoliae (causing black shoot blight) or from Pseudomonas syringae pv. syringae (causing shoot blight on apple trees). The detection limit of DS-LAMP was 10 cells per LAMP reaction, equivalent to $10^4$ cells per ml of the sample extract. DS-LAMP successfully diagnosed the pathogens on four fire-blight infected apple and pear orchards. In addition, it could distinguish black shoot blight from fire blight. The $B{\ddot{u}}hlmann$-LAMP, developed previously for on-site diagnosis of fire blight, did not give consistent results for specificity to E. amylovora and on-site diagnosis; it gave positive reactions to three strains of E. pyrifoliae and two strains of P. syringae pv. syringae. It also, gave positive reactions to some healthy sample extracts. DS-LAMP, developed in this study, would give more accurate on-site diagnosis of fire blight, especially in the Republic of Korea, where fire blight and black shoot blight coexist.

• Asian-Australasian Journal of Animal Sciences
• /
• 제33권10호
• /
• pp.1579-1589
• /
• 2020

Objective: This study was conducted to investigate the roles of LIM kinases (LIMK1 and LIMK2) during porcine early embryo development. We checked the mRNA expression patterns and localization of LIMK1/2 to evaluate their characterization. We further explored the function of LIMK1/2 in developmental competence and their relationship between actin assembly and cell junction integrity, specifically during the first cleavage and compaction. Methods: Pig ovaries were transferred from a local slaughterhouse within 1 h and cumulus oocyte complexes (COCs) were collected. COCs were matured in in vitro maturation medium in a CO2 incubator. Metaphase II oocytes were activated using an Electro Cell Manipulator 2001 and microinjected to insert LIMK1/2 dsRNA into the cytoplasm. To confirm the roles of LIMK1/2 during compaction and subsequent blastocyst formation, we employed a LIMK inhibitor (LIMKi3). Results: LIMK1/2 was localized in cytoplasm in embryos and co-localized with actin in cell-to-cell boundaries after the morula stage. LIMK1/2 knockdown using LIMK1/2 dsRNA significantly decreased the cleavage rate, compared to the control group. Protein levels of E-cadherin and β-catenin, present in adherens junctions, were reduced at the cell-to-cell boundaries in the LIMK1/2 knockdown embryos. Embryos treated with LIMKi3 at the morula stage failed to undergo compaction and could not develop into blastocysts. Actin intensity at the cortical region was considerably reduced in LIMKi3-treated embryos. LIMKi3-induced decrease in cortical actin levels was attributed to the disruption of adherens junction and tight junction assembly. Phosphorylation of cofilin was also reduced in LIMKi3-treated embryos. Conclusion: The above results suggest that LIMK1/2 is crucial for cleavage and compaction through regulation of actin organization and cell junction assembly.

• 한국균학회지
• /
• 제22권4호
• /
• pp.343-349
• /
• 1994

밤나무 동고병(胴枯病)에 심하게 감염(感染)된 미국밤나무의 이병(罹病)줄기로부터 병원균(病原菌)인 Cryphonectria parasitica를 102균주(菌株) 분리(分離)하여 배지상에서 배양(培養)하면서 체세포(體細胞) 화합성(和合性)을 조사한 결과, 102 균주는 54 개의 화합성군(和合性群)으로 나누어 졌으며 그중 38개의 화합성군에 단지 한 균주씩, 6개의 화합성군에 각각 2 균주씩 포함되었으며 나머지 52개 균주는 10개의 화합성군에 포함되어 다양한 화합성군으로 분화(分化)되었음을 보여주었다. 체세포 화합성에 있어서 이러한 다양(多樣)한 분화는 병원균이 분리된 지역에서 80년 이상 존재(存在)해 오면서 시간이 경과(經過)함에 따라 유전적(遺傳的)으로 많은 분화가 일어나고 이로 인하여 화합성군의 숫자가 증가(增加)한 것에 기인(起因)하는 것으로 추측된다. 가장 빈도(頻度)가 높은 10개의 화합성군으로부터 각각 한 균주씩을 대표적인 균주로 선발(選拔)하고 이들을 여러 지역에서 분리된 저병원성 균주와 균사융합(菌絲融合)을 시키면서 저병원성(低病原性) 균주로의 변환(變換)을 시도(試圖)하였다. 10개의 대표적인 균주는 모두 최소(最少)한 1개 이상의 저병원성 균주에 의해서 저병원성 균주로 변환되었다.

• Molecules and Cells
• /
• 제27권1호
• /
• pp.47-54
• /
• 2009

A cDNA clone for a transcript preferentially expressed during an early phase of flooding was isolated from Nicotiana tabacum. Nucleotide sequencing of the cDNA clone identified an open reading frame that has high homology to the previously reported glycine-rich RNA-binding proteins. The open reading frame consists of 157 amino acids with an N-terminal RNA-recognition motif and a C-terminal glycine-rich domain, and thus the cDNA clone was designated as Nicotiana tabaccum glycine-rich RNA-binding protein-1 (NtGRP1). Expression of NtGRP1 was upregulated under flooding stress and also increased, but at much lower levels, under conditions of cold, drought, heat, high salt content, and abscisic acid treatment. RNA homopolymer-binding assay showed that NtGRP1 binds to all the RNA homopolymers tested with a higher affinity to poly r(G) and poly r(A) than to poly r(U) and poly r(C). Nucleic acid-binding assays showed that NtGRP1 binds to ssDNA, dsDNA, and mRNA. NtGRP1 suppressed expression of the fire luciferase gene in vitro, and the suppression of luciferase gene expression could be rescued by addition of oligonucleotides. Collectively, the data suggest NtGRP1 as a negative modulator of gene expression by binding to DNA or RNA in bulk that could be advantageous for plants in a stress condition like flooding.

• The Plant Pathology Journal
• /
• 제21권1호
• /
• pp.13-20
• /
• 2005

Chlorella viruses are large icosahedral, plaque-forming, dsDNA viruses that infect certain unicellular, chlorellalike green algae. The genomic DNA of over 300 kb contains many useful genes and promoters. Over 40 chlorella viruses have been isolated from fresh water in Korea since 1998. The viruses were amplified initially in chlorella strain NC64A, and pure isolates were obtained by repeated plaque isolation. SDS-PAGE analysis revealed similar but distinct protein patterns, both among the group of purified viruses and in comparison with the prototype chlorella virus PBCV-1. Digestions of the 330- to 350-kb genomic DNAs with 10 restriction enzymes revealed different restriction fragment patterns among the isolates. The tRNA-coding regions of 8 chlorella viruses were cloned and sequenced. These viruses contain 14-16 tRNA genes within a 1.2- to 2-kb region, except for the SS-1 isolate, which has a 1039-bp spacer in a cluster of 11 tRNA genes. Promoter regions of several early genes were isolated and their activities were analyzed in transformed chlorella. Some promoters showed stronger activity than commonly used CaMV 35S promoter and chlorella transformation vectors for heterologous protein are beings constructed using these promoters.