• 한국잡초학회지
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• 제30권2호
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• pp.122-134
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• 2010

본 연구의 목적은 다양한 생물종의 Protox 유전자 과다발현 제초제 저항성 형질전환 벼의 저항성 수준과 생육저해 원인이 생육시기별 tetrapyrrole 중간물질 축적과 대사 경로 물질, 활성산소 발생, 지질과산화 작용 및 항산화 효소 능력과 관련성이 있는지를 조사하는데 있다. Myxococcus xanthus(MX, MX1, PX), Arabidopsis thaliana(AP31, AP36, AP37) 및 인간(H45, H48, H49) Protox 과다발현 형질전환벼는 비형질전환벼에 비해 oxyfluorfen에 각각 43~65, 41~72 및 17~70배 저항성을 보였다. 다양한 생물종의 Protox 유전자 과다발현 제초제 저항성 형질전환 벼 중에 일부 라인은 다른 광조건하에서 정상적인 생육을 보인 반면에 일부 라인은 초장 및 생체중 감소가 야기되었다. 그러나 일관성 있게 파종 후 7일과 14일에 초장의 감소가 야기 되었던 형질전환 라인은 AP37뿐이었다. 또한 이들 형질전환 벼 라인들은 저광 및 암조건보다 고광조건하에서 초장 및 생체중 감소가 뚜렷하였다. 파종 후 7일째 Proto IX 축적은 AP31, AP37 및 H48에서 비형질전환벼에 비해 유의적으로 많았고, 파종 후 14일째에는 PX, AP37 및 H48에서 유의적으로 많았다. 파종 후 7일째 Mg-Proto IX은 일부 라인(H41과 H48)을 제외하고 그리고 Mg-Proto IX monomethyl ester는 MX, MX1, PX를 제외한 형질전환 라인 간에 차이가 없었다. 비록 terapyrrole 중간물질이 축적 되었더라도 이들 축적량은 Protox 과다발현 형질전환벼의 생육을 저해 하는데 충분하지 않았을 것으로 생각된다. 또한 활성산소종 ($H_2O_2$와 ${O_2}^{{\cdot}_-}$), MDA, ALA 합성 및 엽록소 함량에서도 형질전환 라인간에 유의적인 차이가 인정되지 않아 tetrapyrrole 축적량이 형질전환 벼의 생육저해에 충분하지 못했음을 확인할 수 있었다. 따라서 일부 형질전환벼에서 초기 생육저해는 어떤 단일 요인에 기인되는 것보다 복합적인 요인에 의해 기인되는 것으로 생각된다.

• Annals of Clinical Microbiology
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• 제22권1호
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• pp.9-13
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• 2019

배경: 다양한 임상검체에서 카바페넴분해효소 생성 장내세균(carbapenemase-producing Enterobacteriace, CPE)의 분리가 증가하고 있다. CPE 감염증은 치사율이 높고, 집단 발병의 가능성이 높아 지속적인 감시가 필수적이다. 저자들은 CPE 주요 유전자들을 한 번에 검출하기 위한 multiplex PCR을 개발하고 이를 평가하고자 하였다. 방법: 총 7종의 내성 유전자를 동시에 검출하기 위한 시발체를 새롭게 디자인하고 PCR 조건을 설정하였다. 주요 카바페넴분해효소인 KPC, IMP, VIM, NDM-1, GES, OXA-23 및 OXA-48 유전자를 대상으로 하였다. 각 유전자의 증폭 산물은 100 bp 이상의 차이가 나도록 고안하였다. 개발된 multiplex PCR은 총 69주의 CPE 양성 임상분리균주를 이용하여 평가하고, 비특이적 증폭에 의한 위양성 가능성의 배제를 위해 71주의 카바페넴 감수성 균주로 확인하였다. 결과: 본 연구에서 개발한 시발체 및 PCR 조건을 이용하여 실험한 결과 CPE 내성 유전자인 KPC 양성 14주, IMP 양성 13주, OXA-23 양성 12주, OXA-48 양성 11주, VIM 양성 9주, GES 및 NDM 양성 각 5주 등 총 69주 모두에서 CPE 유전자의 검출이 가능함을 확인하였다. 카바페넴 감수성 균주를 이용한 특이성 확인 시험에서 Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa 등을 포함한 총 71주의 다양한 그람양성 및 음성균주 모두에서 음성임을 확인하였다. 결론: 본 연구에서 개발된 multiplex PCR은 총 7종의 CPE 유전형을 한 번에 검출할 수 있어 CPE 확인 시험에 유용할 것으로 생각한다.

• 한국작물학회지
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• 제68권3호
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• pp.134-146
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• 2023

콩 역병(Phytophthora root rot, PRR)은 난균(oomycete)인 Phytophthora sojae에 의해 발생하는 콩의 주요 병 중 하나로, 배수가 잘 안 되는 밭이나 습한 토양에서 심하게 발생한다. 역병의 피해를 효과적으로 줄일 수 있는 방법은 주로 역병 저항성 품종을 재배하는 것으로, 이는 저항성 유전자 Rps (resistance to P. sojae)에 대한 연구를 중심으로 이루어진다. 본 연구는 대풍 과 천알(계통명 SS0404-T5-76)을 교배하여 구축한 RIL (recombinant inbred line) 집단을 이용하여 콩 역병 균주40468과 연관된 저항성 유전자좌를 탐색하기 위해 수행되었다. 역병 균주40468에 대한 저항성 평가는 하배축 접종(hypocotyl inoculation) 방법으로 이루어졌다. 저항성 검정 결과, 천알은 저항성,대풍은 감수성을 보였고 집단 내에서는 계통들의 표현형이 분리되는 양상을 보였다. 집단 내에서 표현형 분포는 1:1 (R:S) (χ2 = 0.57, p = 0.75) 분리비와 일치하였으며, 이는 저항성 반응이 단일 유전자에 의해 조절됨을 나타낸다. 대풍, 천알과 각 RIL 계통들은 고밀도 SNP 유전자형 분석을 통해 데이터를 얻었고, 이를 바탕으로 유전자 지도를 작성하였다. 일원분산 분석(Single-marker ANOVA) 및 linkage analysis 결과, 18번 염색체의 55.9~56.4 Mbp에서 높은 통계적 유의성을 보였으며, 이 지역의 표현형 분산은 ~98%로 나타났다. 탐색된 영역은 다수의 선행연구에서 Rps의 위치로 보고된 지역과 겹치며, 콩 표준 유전체 정보를 기반으로 0.5 Mbp 범위 내에서 leucine-rich repeat (LRR) 또는 serine/threonine kinase(STK)을 합성하는 유전자 9개를 포함하고 있다. 천알은 역병 균주40468에 대한 저항성 유전자좌가 밝혀진 첫 국내 콩 품종으로, 본 연구에서 밝힌 천알의 저항성 유전자좌는 향후 역병 저항성 육종 및 연구에서 유용한 재료가 될 것이다.

• Journal of Microbiology
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• 제44권6호
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• pp.660-664
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• 2006

The relationship between H. pylori clarithromycin resistance and genetic pattern distribution has been differently explained from different geographic areas. Therefore, we aimed to assess the clarithromycin resistance rate, to evaluate the bacterial genetic pattern, and to search for a possible association between clarithromycin resistance and cagA or vacA genes. This prospective study enrolled 62 consecutive H. pylori infected patients. The infection was established by histology and rapid urease test. Clarithromycin resistance, cagA and vacA status, including s/m subtypes, were assessed on paraffin-embedded antral biopsy specimens by TaqMan real time polymerase chain reaction (PCR). Primary clarithromycin resistance was detected in 24.1 % of cases. The prevalence of cagA was 69.3%, and a single vacA mosaicism was observed in 95.1 % cases. In detail, the s1m1 was observed in 23 (38.9%) patients, the s1m2 in 22 (37.2%), and the s2m2 in 14 (23.7%), whereas the s2m1 combination was never found. The prevalence of cagA and the vacA alleles distribution did not significantly differ between susceptible and resistant strains. Primary clarithromycin resistance is high in our area. The s1m1 and s1m2 are the most frequent vacA mosaicisms. There is no a relationship between clarithromycin resistance and bacterial genotypic pattern and/or cagA positivity.

• BMB Reports
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• 제40권5호
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• pp.773-782
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• 2007

Different isolates of the soil bacterium Bacillus thuringiensis produce multiple crystal (Cry) proteins toxic to a variety of insects, nematodes and protozoans. These insecticidal Cry toxins are known to be active against specific insect orders, being harmless to mammals, birds, amphibians, and reptiles. Due to these characteristics, genes encoding several Cry toxins have been engineered in order to be expressed by a variety of crop plants to control insectpests. The cotton boll weevil, Anthonomus grandis, and the fall armyworm, Spodoptera frugiperda, are the major economically devastating pests of cotton crop in Brazil, causing severe losses, mainly due to their endophytic habit, which results in damages to the cotton boll and floral bud structures. A cry1Ia-type gene, designated cry1Ia12, was isolated and cloned from the Bt S811 strain. Nucleotide sequencing of the cry1Ia12 gene revealed an open reading frame of 2160 bp, encoding a protein of 719 amino acid residues in length, with a predicted molecular mass of 81 kDa. The amino acid sequence of Cry1Ia12 is 99% identical to the known Cry1Ia proteins and differs from them only in one or two amino acid residues positioned along the three domains involved in the insecticidal activity of the toxin. The recombinant Cry1Ia12 protein, corresponding to the cry1Ia12 gene expressed in Escherichia coli cells, showed moderate toxicity towards first instar larvae of both cotton boll weevil and fall armyworm. The highest concentration of the recombinant Cry1Ia12 tested to achieve the maximum toxicities against cotton boll weevil larvae and fall armyworm larvae were 230 ${\mu}g/mL$ and 5 ${\mu}g/mL$, respectively. The herein demonstrated insecticidal activity of the recombinant Cry1Ia12 toxin against cotton boll weevil and fall armyworm larvae opens promising perspectives for the genetic engineering of cotton crop resistant to both these devastating pests in Brazil.

• Asian-Australasian Journal of Animal Sciences
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• 제21권4호
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• pp.489-493
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• 2008

In 2004, Jorgensen and coworkers proposed the MUC4 gene as a candidate gene of enterotoxigenic Escherichia coli (ETEC) F4ab/ac receptor in piglets and a mutation of $G{\rightarrow}C$ in intron 7 of MUC4 was identified to be associated with the ETEC F4ab/ac adhesion phenotypes. In this study, we used 310 piglets of three breeds (Landrace, Large White and Chinese Songliao Black) to analyze the relationship between this mutation and the F4ab/ac adhesion phenotype. The results show that the genotypes at this site and the ETEC F4ab/ac adhesion phenotypes were not completely consistent, although they are very strongly associated. Among the individuals with genotype CC, which was identified as a resistant genotype to F4ab/ac adhesion, only 72.1% (124/172) were non-adhesive to ETEC F4ab and 77.9% (134/172) were non-adhesive to ETEC F4ac infections. This suggests that this mutation may not be the causative mutation for ETEC F4ab/ac adhesion, rather, the actual causative mutation may be in another gene closely linked to MUC4, or at aother site within the MUC4 gene. Our results also suggest that the receptors of F4ab and F4ac may be determined by two different but closely linked loci. In order to screen other genes related to F4ab/ac adhesion in piglets, the mRNA profiles from six full sib piglets, of which three were adhesive to ETEC F4ab/ac and three non-adhesive, were analyzed by suppression subtractive hybridization (SSH). One up-regulated gene, Ep-CAM, was selected for further analysis based on its role in the intestinal epithelial cells adhesion. Using real-time RT-PCR, we found that the Ep-CAM gene was significantly up-regulated in the piglets adhesive to F4ab/ac. It was mapped to SSC3q11-q14 by radiation hybrid mapping.

• IMMUNE NETWORK
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• 제1권1호
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• pp.45-52
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• 2001

Background: Many types of cancer become resistant to current chemotherapeutic and radiotherapeutic intervention. To overcome this situation application of gene therapy by the introduction of suicide genes followed by their prodrugs may be promising. A viral enzyme, Herpes simplex thymidine kinase (HSV-tk), which converts ganciclovir from an inactive prodrug to a cytotoxic agent by phosphorylation, are being actively investigated for use in gene therapy for cancer. The purpose of this study was to determine whether combining prodrug-activating gene therapy and irradiation might result in enhanced antitumor effects. Methods: The HSV-tk gene was cloned into the retroviral vector, pLXSN and established the clones producing retroviruses carrying the HSV-tk gene. The carcinoma cell line, HCT116 and Huh-7 were transduced with high-titer recombinant retroviruses. These cell lines were treated with ganciclovir before or after irradiation for the defining combinational effect of suicide gene therapy and radiotherapy. Results: The titers of cloned PA3 17 amphotropic retroviruses ranged from 4 to 6 X $10^6CFU/ml4$. After selectional periods, the expression of HSV-tk was confirmed by reverse-transcriptase polymerase chain reaction (RT-PCR). The growth of cells expressing HSV-tk was inhibited as increase of GCV dose after 48 hr and the growth inhibitory effect of GCV was much higher after 72 hr. When the cells transduced with HSV-tk gene were exposed to radiation, the growth inhibitory effect of GCV was significantly increased, as compared with non-transduced parental cells. Conclusions: The results suggest that the addition of HSV-tk gene therapy to standard radiation therapy may improve the effectiveness of treatment for solid tumors.

• Korean Journal of Acupuncture
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• 제37권2호
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• pp.63-75
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• 2020

Objectives : Osteoporosis is the most common bone disease and osteoporosis fracture is the leading cause of decreased life. Bisphosphonate and selective estrogen receptor modulators are the best choice of treatment for osteoporosis. However, when used for a long time, they increase the probability of side effect such as osteonecrosis of the jaw. Thus, it is crucial to develop alternative medicine to treat osteoporosis. Gentianae Macrophyllae Radix, a herbal medicine, is mainly to treat rheumatoid arthritis. However, the effect of the water extract of Gentianae Macrophyllae Radix (w-GM) on osteoporosis has not been investigated. Thus, we examine whether w-GM can inhibit osteoclast differentiation and bone resorption on receptor activator of nuclear factor kappa-B (NF-κB) ligand (RANKL)-treated RAW 264.7 cells. In this study, RAW 264.7 cells were used as an osteoclast differentiation model by treating them with RANKL. Methods : RAW 264.7 cells were used to determine the effect of w-GM on osteoclast differentiation and bone resorption. The number of tartrate-resistant acid phosphatase (TRAP)-positive cells, TRAP activity and pit formation assay were examined. In addition, protein expressions were measured by western blot and mRNA expressions were analyzed by reverse transcription polymerase chain reaction. Results : Treatment with w-GM inhibited the number of TRAP-positive cells, TRAP activity and pit area. In addition, w-GM decreased protein expression such as mitogen-activated protein kinase, NF-κB, c-Fos and nuclear factor of activated T-cells cytoplasmic 1 (NFATc1). It also inhibited the mRNA levels such as c-Fos, NFATc1, TRAP, NF-κB, calcitonin receptor and cathepsin K in RANKL-treated RAW 264.7 cells. Conclusions : These results suggest that w-GM has inhibitory effects via osteoclast differentiation, thus it could be a new medication for osteoporosis.

• 한국미생물·생명공학회지
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• 제37권2호
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• pp.110-117
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• 2009

독도에 자생하고 있는 야생 가지과 식물근권세균들의 특성을 밝히기 위해 까마중을 채집하였다. 까마중 근권으로부터 총 44종의 포자형성세균 혹은 질소고정세균을 분리하였다. 이 균주들은 16S rDNA 염기서열을 이용하여 부분 동정하였다. PGPR로써의 특성을 밝히기 위해 옥신 생산능, 인 가용능, 그리고 Siderophore 형성능을 측정하였다. 식물 생장호르몬인 옥신을 형성하는 균주는 19종, 난분해성 인을 분해할 수 있는 균주는 8종, 식물병원균을 억제하는 siderophore를 형성할 수 있는 균주는 13종으로 확인되었다. 우리나라에서 널리 재배되고 있는 가지과식물인 담배와 고추에 직접 적용하여, 식물의 생장 촉진 효과 및 식물 전신유도저항성 효과를 확인하였다. 특히 KUDC1009는 식물의 생장 촉진 효과 그리고 식물병원균에 대한 저항성을 증가시키는 등 다기능의 능력을 가지는 것을 확인할 수 있었다. 독도의 근권세균들은 다양한 스트레스 환경하에서 야생 까마중이 생존하는데 많은 도움을 주는 것을 추측 할 수 있었다.

• Journal of Nutrition and Health
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• 제37권9호
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• pp.786-793
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• 2004

Women with menopause or rats with ovariectomy is associated with increased body weight, body fat and insulin resistance, which are components of metabolic syndrome. Increased prevalence of metabolic syndrome after menopause might be associated with mitochondrial dysfunction, since mitochondrial oxidative and phosphorylation activity is strongly correlated with insulin sensitivity. Although estradiol replacement prevents the metabolic syndrome, harmful effect of estradiol hampers the casual usage to prevent the metabolic syndrome. It has been reported that genistein has a mild estrogenic activity, decreases fat mass in mice and has an antidiabetic role in diabetic rats. Although insulin resistance is closely related to mitochondrial functions, there has not been yet any study in regard to the effect of dietary genistein on mitochondrial function in the insulin resistant female subjects induced by ovariectomy or similar situation. The present study investigated whether the supplementation of genistein in the high fat diet affected the mitochondrial function of high fat fed ovariectomized rats. Female Sprague Dawley rats (8 weeks old) were assigned to the following groups: sham-operated+ high fat diet (S, n=6); sham-operated + high fat diet with 0.1% genistein (S + G, n=7); ovariectomized + high fat diet (OVX, n=8); ovariectomized + high fat diet with 0.1% genistein (OVX+ G, n=8). Ovariectomy significantly increased body weight compared with S group. Genistein consumption in ovariectomized (OVX + G) rats decreased body weight gain compared with OVX rats. Liver weights were increased by ovariectomy. The hepatic mitochondrial protein density expressed as mg per g liver was lower in the OVX group than in the S group. However, OVX + G group showed the increased mitochondrial protein density similar to the level of S group. When mRNA levels of genes related to mitochondria such as peroxisome proliferator-activated receptor ${\gamma}$ coactivator 1 (PGC-1) and cytochrome c oxidase subunit III (COX III) were measured, there were decreases in the mRNA levels of PGC-1 and COX III in S + G, OVX and OVX + G group. The activity of cytochrome c oxidase was not different between groups. We could observe the decrease in succinate dehydrogenase (SDH) activity per g liver in OVX rats. Genistein supplement increased SDH activity. In conclusion, genistein supplementation to the OVX rats enhanced mitochondrial function by increasing mitochondrial protein density and SDH activity. The improvement in mitochondrial function by genistein can contribute to the improvement in metabolic syndrome.