• The Plant Pathology Journal
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• 제26권4호
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• pp.386-393
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• 2010

In order to study disease resistance mechanisms in rice against the rice blast fungus Magnaporthe grisea, we screened fungal elicitor-responsive genes from rice suspension-cultured cells treated with fungal elicitors employing differential hybridization (DH). By DH screening, 31 distinct rice clones were isolated and a majority of them were full-length cDNAs encoding pathogenesisrelated (PR) genes. Sixteen of the 31 genes were upregulated at 4, 8, and 12 h following fungal elicitor treatment. To elucidate the effect of signal molecules and biotic elicitors on the regulation of rice defense genes, we further characterized the transcriptional expression patterns of representative isolated PR genes; OsGlu1, OsGlu2, OsTLP, OsRLK, and OsPR-10, following treatment with fungal elicitor, phytohormones, cycloheximide, and inhibitors of protein phosphorylation. Jasmonic acid (JA) induced transcriptional expression of OsGlu1, OsTLP, and OsRLK, but not of OsGlu2 and OsPR-10 at any of the tested time points. Salicylic acid (SA) and abscisic acid weakly induced the expression of OsTLP and OsRLK. SA showed an antagonistic effect with fungal elicitor and JA. Cycloheximide suppressed all these genes upon elicitor treatment, except for OsGlu2. Staurosporine only induced the expression of OsRLK. Application of calyculin A strongly induced OsRLK expression, but suppressed the expression of OsGlu2. Our study yielded a number of PR genes that play a role in defense mechanisms against the rice blast fungus, as well as contribute towards the elucidation of crosstalk between phytohormones and other modifications during defense signaling.

• Radiation Oncology Journal
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• 제19권4호
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• pp.389-396
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• 2001

목적 : 임상에서 사용하는 방사선량을 조사하여 환자의 자궁경부암 세포에서 유도되는 유전자를 검색하고자 하였다. 대상 및 방법 : 자궁경부암 환자에서 방사선치료 하루 전(대조군)과 1.8 Gy 조사후 40분 지나서(조사군) 자궁경부암 조직을 생검하여 각 군에서 total RNA를 추출하였다. differential display reverse transcription-polymerase chain reaction기법(DDRT-PCR)으로 발현이 증가 또는 감소된 유전자를 탐색하였다. 발현에 변화가 있는 cDNA를 추출하고 증폭하여 얻은 클론을 reverse Northern Blot방법을 이용하여 screening하였고, Northern Blot으로 확인하였다. sequencing을 실시한 후 NCBI database를 이용하여 blast search를 하였다. 발현이 감소한 유전자를 대상으로 다른 환자에서의 발현 양상을 확인하기 위하여 방사선치료를 받는 5명의 자궁경부암 환자를 대상으로 RT-PCR로 검사 하였다. 결과 : DDRT-PCR기법을 이용하여 방사선 조사군에서 발현이 증가 혹은 감소된 18개의 cDNA band를 발견하였다. reverse northern blot을 이용한 screening에서 발현이 증가된 10개의 클론과 감소된 1개의 클론을 확인하였다. 클로닝된 cDNA 조각들은 대부분 $400\~500\;bp$ 정도 되었으며, 그중 1개의 클론은 잘 알려져 있는 chemokine receptor CXCR4 유전자와 높은 상동성을 보였으며, 4개의 클론은 Human ESTs로 확인되었고 5개의 클론은 기능이 아직 확인되지 않은 알려져 있는 염기서열로 확인되었다. 방사선에 의하여 발현이 감소한 CxCa-11 클론이 모든 환자에서 방사선치료 전 시료에서 발현을 보였으나, 방사선치료 후 시료에서는 그 발현량이 감소하거나 발현이 안되었다. 결론 : DDRT-PCR을 이용하여 임상에서 자주 사용되는 방사선량을 환자의 자궁경부암에 조사했을 때 발현되는 유전자를 확인하였다. 이러한 유전자 발현의 확인은 방사선치료과정 중에 발생하는 일련의 기전들을 이해하는데 도움이 되리라 생각된다.

• Pediatric Infection and Vaccine
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• 제21권1호
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• pp.65-70
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• 2014

선천성 매독은 산전진찰로 예방할 수 있음에도 불구하고, 여전히 문제가 되고 있다. 최근에는 선천성 매독과 관련된 혈뇨나 폐렴이 보고된 증례가 없었으나 저자들은 적절한 산전치료를 받지 못한 산모에게서 태어난 22일된 남아에게서 발생한 선천성 매독에 동반된 신증후군과 폐렴의 증례를 보고한다. 선천 매독의 진단은 혈청학적 검사로 확진하였고, 환아는 페니실린 G 치료를 받고 회복되었다. 신생아에게서 임상증상은 매우 불확실할 수 있어, 질환에 대한 인식이 지연 될 수 있다. 따라서 선천성 매독의 예방에는 철저한 산전관리가 필수적이다. 만약 산모가 적절한 산전진찰을 받지 않았을 경우에는 신속한 혈청학적 진단이 반드시 시행되어야 할 것이다.

• Molecules and Cells
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• 제39권5호
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• pp.426-438
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• 2016

Plant disease resistance occurs as a hypersensitive response (HR) at the site of attempted pathogen invasion. This specific event is initiated in response to recognition of pathogen-associated molecular pattern (PAMP) and subsequent PAMP-triggered immunity (PTI) and effector-triggered immunity (ETI). Both PTI and ETI mechanisms are tightly connected with reactive oxygen species (ROS) production and disease resistance that involves distinct biphasic ROS production as one of its pivotal plant immune responses. This unique oxidative burst is strongly dependent on the resistant cultivars because a monophasic ROS burst is a hallmark of the susceptible cultivars. However, the cause of the differential ROS burst remains unknown. In the study here, we revealed the plausible underlying mechanism of the differential ROS burst through functional understanding of the Magnaporthe oryzae (M. oryzae) AVR effector, AVR-Pii. We performed yeast two-hybrid (Y2H) screening using AVR-Pii as bait and isolated rice NADP-malic enzyme2 (Os-NADP-ME2) as the rice target protein. To our surprise, deletion of the rice Os-NADP-ME2 gene in a resistant rice cultivar disrupted innate immunity against the rice blast fungus. Malic enzyme activity and inhibition studies demonstrated that AVR-Pii proteins specifically inhibit in vitro NADP-ME activity. Overall, we demonstrate that rice blast fungus, M. oryzae attenuates the host ROS burst via AVR-Pii-mediated inhibition of Os-NADP-ME2, which is indispensable in ROS metabolism for the innate immunity of rice. This characterization of the regulation of the host oxidative burst will help to elucidate how the products of AVR genes function associated with virulence of the pathogen.

• 한국잡초학회지
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• 제16권3호
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• pp.187-193
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• 1996

금강아지풀, 가을강아지풀, 강아지풀 종자의 발아특성과 제초제에 대한 반응차이를 조사하여 시험재료로서 발아력이 양호한 종자를 효율적으로 공급할 수 있는 조건들을 찾음은 물론 종간 발아특성 차이와 제초제에 대한 반응차이간의 관련성 여부를 검토하여 보았다. 금강아지풀의 경우 실온건조에서, 가을강아지풀 및 강아지풀의 경우는 저온습윤에서 가장 빨리 휴면타파 되었다. 휴면의 정도는 금강아지풀, 강아지풀, 가을강아지풀 순으로 강한 경향이었다. 금강아지풀의 경우 휴면타파된 종자를 저온습윤 또는 저온건조에 보관하더라도 발아력에 변함이 없었으나 가을강아지풀 및 강아지풀의 경우는 저온건조에 두면 발아속도가 늦어지며 초기발아율이 떨어지는 경향이었으나 50% 이상의 발아력(2개월째)은 유지하였다. 금강아지풀 종자를 실온건조에 보관할 경우 6년 이상이면 발아력이 감퇴되었다. 발아는 세 종류 모두 $30^{\circ}C$ 14시간/$20^{\circ}C$ 10시간의 온도조건에서 양호하였으며 광의 유무에 따른 발아율 타이는 인정되지 않았다. Cycloxydim, sethoxydim, primisulfuron 처리에서는 종간 반응차이가 없었으나, fenoxapropethyl은 가을강아지풀과 강아지풀에서, fluazifopbutyl은 가을강아지풀에서, clorimuron-ethyl은 금강아지풀파 가을강아지풀에서 감수성 정도가 약간 높았을 뿐이었다. 반면에 EK-2612는 반응 차이가 현저하여 강아지풀보다 금강아지풀에서 높은 감수성을 보였다. 이로 볼 때 동일 작용메카니즘을 가지는 약제들간에도 종간에 일관된 반응차이를 가지는 것이 아니라 오히려 화합물에 의존하는 반응을 보였으며, 종간 발아특성 차이와 제초반응과는 어떠한 관련성이 없었다.

• 한국응용곤충학회지
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• 제35권4호
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• pp.321-325
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• 1996

누에에서 새로운 항세균성 펩타이드 유전자를 탐색하기 위하여 E. coli K12로 체강 주사한 누에 유충의 cDNA 유전자 은행에서 차별화 선별로, 잠재 항세균성 펩타이드 유전자로 추정되는 BmInc8 클론을 분리하였다. BmInc8은 564bp의 크기를 가지며, 59개 아미노산을 coding하는 open reading frame과 2개의 잠정 폴리아데닐화 부위를 보유하고 있었다. BmInc8은 M. sexta에서 분리, 보고된 bactericidine 유전자와 61.2%의 DNA 상동성을 나타내는 것으로, 그 연역된 펩타이드 구조는 항세균성 펩타이드의 일종인 cecropin과 유사한 2가닥의 $\alpha$-helix가 Lysine-Proline 경첩부위에 의해 포개져 있는 형태를 갖는 것으로 추정되었다. 또한 cDNA 삽입 부위의 기능성 검정을 위해 원핵 발현벡터인 pT7-5를 이용하여 E. coli BL21(DE3) 균주에 형질전환하고 IPTG로 induction한 결과 E. coli BL21(DE3) 균주의 성장이 정지됨을 관찰할 수 있었다. 이상의 결과로 BmInc8은 DNA 상동성 비교, 연역 아미노산의 구조 추정 및 cDNA 삽입 부위를 이용한 transient expression 결과 항세균성 펩타이드를 coding하는 유전자임을 추정할 수 있었다. 또한 Bminc8의 cDNA 유전자 정보를 GenBank에 등록하였으며 등록 번호는 U30289이었다.

• BMB Reports
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• 제33권5호
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• pp.396-401
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• 2000

A differential display method is used to identify novel genes whose expression is affected by treatment with ferric ammonium citrate (FAC) or desferrioxamine (DFO), an iron chelating agent in the human hepatoblastoma cell line (HepG2). These chemicals are known to deplete or increase the intracellular concentration of iron, respectively. Initially, we isolated seventeen genes whose expressions are down- or up regulated by the treatment of the chemicals, as well as their four differentially expressed genes that are designated as clone-1, -2, -3, and -4. These are further characterized by cDNA sequencing and Northern blot analysis. Through the cDNA sequencing, as well as comparing them to genes published using the NCBI BLAST program, we identified the sequence of the clone-1 that is up-regulated by the treatment of DFO. It is identical to the human insulin-like growth factor binding protein-1 (IGFBP-1). This suggests that the IGFBP-1 gene in the HepG2 cell is up-regulated by an iron depletion condition. Also, the expression of the clone-3 and -4 is up-regulated by FAC treatment and their eDNA sequences are identical to the human ferritin-fight chain and human NADH-dehydrogenase, respectively. However, the sequence of the clone-2 has no significant homology to any other known gene. Therefore, we suggest that changes of the cellular iron level in the HepG2 cell affects the transcription of cellular genes. This includes human IGFBP-1, ferritin-fight chain, and NADH-dehydrogenase. Regulation of these gene expressions may have an important role in cellular functions that are related to cellular iron metabolism.

• 한국발생생물학회지:발생과생식
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• 제16권4호
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• pp.301-307
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• 2012

Olive flounder Paralichthys olivaceus is one of the most widely cultured fish species in Korea. Although olive flounder receive attention from aquaculture and fisheries and extensive research has been conducted eye morphological change in metamorphosis, but little information was known to molecular mechanism and gene expression of eye development- related genes during the early part of eye formation period. For the reason of eyesight is the most important sense in flounder larvae to search prey, the screening and identification of expressed genes in the eye will provide useful insight into the molecular regulation mechanism of eye development in olive flounder. Through the search of an olive flounder DNA database of expressed sequence tags (EST), we found a partial sequence that was similar to crystallin beta A1 and gamma S. Microscopic observation of retinal formation correspond with the time of expression of the crystallin beta A1 and gamma S gene in the developmental stage, these result suggesting that beta A1 and gamma S play a vital role in the remodeling of the retina during eye development. The expression of crystallin beta A1 and gamma S were obviously strong in eye at all tested developing stage, it is also hypothesized that crystallin acts as a molecular chaperone to prevent protein aggregation during maturation and aging in the eye.

• 원예과학기술지
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• 제31권3호
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• pp.275-281
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• 2013

Low temperature is one of the major environmental factors that affect productivity including reduced growth and budding of vines, and changes of metabolic processes in grape (Vitis spp.). To screen the specific expression of abiotic stress-related genes against cold treatment in 'Kyoho' and 'Campbell Early' grapevines, expression of various defense-related genes was investigated by RT-PCR and real-time PCR. Among the 67 genes analyzed by RT-PCR and real-time PCR, 17 and 16 types of cDNA were up-regulated, while 5 and 6 types were down-regulated in cold-treated 'Kyoho' and 'Campbell Early' grapevines, respectively. Genes encoding carotene (Cart3564 and Cart4472), chalcone isomerase (CHI), cytochrome P450 (CYP), flavonol synthase (FLS), endo-${\beta}$-glucanase precursor (Glu), glutathione peroxidase (GPX), glutathione-S-transferase (GST), leucine-rich repeats (LRR), manganese superoxide dismutase (Mn-SOD), phenylalanine ammonia lyase (PAL), polygalacturonase-inhibiting protein (PGIP), proline rich protein 2 (PRP2), small heat shock protein (sHSP), temperature induced lipocalin (TIL), and thaumatin-like protein (TLP) were up-regulated, while those encoding CBF like transcription factor (CBF1), chitinase-like protein (CLP), cold induced protein (CIP), glycerol-3-phosphate acyltransferase (GPAT), and mitogen-activated protein kinase (MAPK) were down-regulated by low temperature treatment in both in 'Kyoho' and 'Campbell Early'.

• 대한약침학회지
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• 제8권1호
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• pp.31-40
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• 2005

Objectives : It has long been known about the osteogenic effect of CTF-HAS on bone tissues. However, it has not been determined the effect of CTF-HAS on cancer cells. The purpose of this study is to screen the CTF-HAS mediated differentially expressed genes in cancer cells such as SNU484 gastric cancer cell lines. Oligonucleotide microarray approach were employed to screen the differential expression genes. Methods : CTF-HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. Cells were treated with various concentrations of CTF-HAS(0.1, 0.5, 1.5, 10, 20mg/ml) for 24 h. Cytotoxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with 1.5mg/ml of CTF-HAS. For oligonucleotide microarry assay, total RNA was used for gene expression analysis using oligonucleotide genechip (Human genome U133 Plus 2.0., Affimatrix Co.). Results : It has no cytotoxic effects on HepG2 cells in all concentration (0.1, 0.5, 1.5, 10,20mg/ml). More than twofold up-regulated genes were 5 genes. The number of more than twofold down-regulated genes was 10. Discussion : This study showed the screening of CTF-HAS mediated differentially regulated genes using combined approaches of oligonucleotide microarray. The screened genes will be used for the better understanding in therapeutic effect of CTF-HAS on cancer field.