• Journal of Plant Biology
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• 제37권3호
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• pp.371-380
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• 1994

To determine the patterns and the levels of expression of the cauliflower mosaic virus (CaMV 35S) promoter and the rice actin 1 (Act1) promoter in rice, transgenic rice plants containing CaMV 35S-$\beta$-glucuronidase (GUS) and Act1-GUS constructs were generated and examined by fluorometric and histochemical analyses. The fluorometric analysis of stably transformed calluses showed that the activity of the rice Act1 promoter was stronger than that of the CaMV 35S promoter in rice cells. In a histochemcial study of the transgenic rices, it was shown that the GUS activity directed by the CaMV 35S promoter was localized mainly in parenchymal cells of vascular tissues of leaves and roots and mesophyll cells of leaves. These results are similar to those of potato, a dicot plant. In contrast, rice plant transformed with Act1-GUS fusion construct revealed strong GUS activity in parenchymal cells of vascular tissue, mesophyll cells, epidermal cells, bulliform cells, guard subsidiary cells of leaves and most cells of the root, suggesting that the rice Act1 promoter is more constitutive than the CaMV 35S promoter. It was also confirmed that in both types of transgenic rice little or no staining was localized in metaxylen tracheary elements of vascular tissue from leaves or roots. These results indicate that the rice Act1 promoter can be utilized more successfully for expression of a variety of foreign gene in rice than the CaMV 35S promoter.

• The Plant Pathology Journal
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• 제18권4호
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• pp.192-198
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• 2002

Mixed infections of two economically important viruses, Turnip mosaic virus(TuMV) in the family Potyviridae and Ribgrass mosaic virus(RMV) in the genus Tobamo-virus, were studied ultrastructurally on oriental cabbage. TuMV-ACl8 (alpine isolate in Korea) induced chlorotic spots on inoculated leaves of both ‘SSD63’ inbred line known as susceptible to TuMV, and ‘Tambok’ commercial cultivar, known as resistant to the virus, in the early stages of infection. TuMV-C5 (Taiwan isolate) caused severe mosaic and malformation on the upper leaves of ‘SSD63’, and necrotic spots in both inoculated and upper leaves of ‘Tambok’. RMV-CA1 (oriental cabbage isolate from alpine in Korea) induced vein chlorosis, leaf malformation, and midrib necrotic streak in the upper leaves of both ‘SSD63’ and ‘Tambok’. Both oriental cabbages infected with a combination of TuMV-ACl8 and RMV-CA1 showed synergistic symptoms of severe yellowing, severe mosaic, and necrotic spot or vein necrosis on their leaves. A combination of TuMV-C5 and RMV-CA1 produced synergistic symptoms only in ‘SSD63’. In ‘Tambok’ infected with the combination of TuMV-C5 and RMV-CA1, the number of necrotic spots on the inoculated leaves was one half lesser than that on singly infected with TuMV-C5. A few necrotic spots progressed systemically. In cells infected with a combination of TuMV-ACl8 and RMV-CA1 or TuMV-C5 and RMV-CA1, the particles of the two viruses made nonagon-like rings(NLR); one TuMV particle was surrounded loosely by nine RMV particles. Two unrelated viruses of TuMV and RMV were compacted in the central part of the spiral aggregates(SA) that was induced strikingly in cells by the mixed infections. The SA showed NLR in its center of the cross-sectioned side. Many particles of RMV of Tobamovirus were closely associated with Potyvirus-characteristic cylindrical inclusions. The SAs in the mixed infections were formed easily by the Potyvirus of TuMV-ACl8 or -C5 isolates.

• KSBB Journal
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• 제7권3호
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• pp.155-160
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• 1992

식량으로 쓰이는 식물 단백질은 공통적으로 Isoleucine, Lysine, Methionine, Threonine, Tryptoplan등 5가지 필수아미노산이 결핍되어있다. 본 연구에서는 이러한 필수아미노산을 다량 함유한 단백질을 발현시킬 수 있는 합성유전자를 fekaqo에서 높은 수준으로 발현시키고자 강한 식물 promoter로 알려진 CaMV 35S, CaMV duplicate 35S promoters를 사용하였다. 형질 전환 및 재분화된 식물을 분석한 결과 본 합성 유전자가 식물 nuclear genome 안으로 도입을 안정하여 잘되었고 mRNA수준까지는 유의적인 증가를 보였으나 단백질 수준에서는 유의적 수준의 증가를 관찰할 수 없었다.

• The Plant Pathology Journal
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• 제19권2호
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• pp.111-116
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• 2003

Turnip mosaic Potyvirus (TuMV) and Ribgrass mosaic Tobamovirus (RMV) are major viruses infecting crucifer crops in Korea. RMV-FG22 was isolated from oriental cabbage. TuMV isolates were TuMV-CA7 from oriental cabbage, TuMV-TU and TuMV-TU2 from turnip, TuMV-RA from rape, TuMV-ST from stock, and TuMV-R9 from radish. The six isolates of TuMV were classified by symptom expression in inbred lines of crucifers. TuMV-CA7 and TuMV-TU isolates infected mostly oriental cabbages; TuMV-ST, TuMV-TU2, and TuMV-R9 infected radishes; and TuMV-RA infected both oriental cabbages and radishes. Crops used in six combinations of mixed infections were 'Tambok' cultivar resistant to TuMV,'SSD63' susceptible inbred line of oriental cabbage, pure line of leaf mustard, and‘Daeburyungyeorum’cultivar of radish. External symptoms in 'Tambok' and radish by each of the six single infections of TuMV showed similar results by bioassay. Synergistic response of necrotic death occurred within 1 week after inoculation in all combinations mixed with TuMV and RMV-FG22 on leaf mustard. In oriental cabbage 'SSD63' , synergism of necrosis occurred in four TuMV isolates, but not in TuMV-ST and TuMV-R9. In oriental cabbage 'Tambok' , synergism was expressed only in two combinations of RMV-FG22+TuMV-CA7 and RMV-FG22+TuV-TU, but other combinations had the same symptoms produced by RM-FG22. In radish‘Daeburyungyeorum’, only mild mosaic symptoms were induced by combinations of RMV-FG22+TuMV-CA7, RMV-FG22+TuMV-TU, RMV-FG22+TuMV-RA, and RMV-FG22+TuMV-R9. Mosaic and severe mosaic were induced in combinations of RMV-FG22 +TuMV-TU2 and RMV-FG22+TuMV-ST, respectively.

• The Plant Pathology Journal
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• 제19권2호
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• pp.117-122
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• 2003

Six isolates of Turnip mosaic Potyvirus (TuMV) namely, TuMV-CA7 from oriental cabbage, TuMV-TU and TuMV-TU2 from turnip, TuMV-RA from rape, TUMV-ST from stock, and TuMV-R9 from radish, and Ribgrass mosaic Tobamovirus (RMV-FG22) from oriental cabbage were isolated. Three kinds of characteristics of the six TuMV isolates were sorted by bioassay: TuMV-CA7 and TuMV-TU isolates infected mostly oriental cabbages; TuMV-ST, TuMV-TU2, and TuMV-R9 infected radishes; and TuMV-RA infected both oriental cabbages and radishes. Mixed infections of crucifers were RMV-FG22+TuMV-CA7, RMV-FG22+TuMV-TU, RMV-FG22+TuMV-RA, RMV-FG22+TuMV-ST, RMV-FG22 +TuMV-TU2 and RMV-FG22+TuMV-R9. Crops used were 'Tambok' cultivar resistant to TuMV, 'SSD63' susceptible inbred line of oriental cabbage, pure line of leaf mustard and 'Daeburyungyeorum' cultivar of radish. New specific ultrastructures of nonagon-like ring (NLR) and spiral aggregates (SA) by mixed infection with TuMV and RMV were formed in cells of crucifer plants. The NLR was made by a TuMV surrounded loosely by nine RMV particles, and the SA was formed spirally by full mixed of two virus particles. The SA had some NLR in its center, which was observed from cross sectioned SA. Host plants with specific ultrastructures expressed synergistic symptoms. Specific ultrastructures of NLR and SA were formed in combinations of RMV-FG22 and in TuMV-CA7, TuMV-TU, or TuMV-RA that could infect oriental cabbages. How-ever, no specific ultrastructures and mixing of the two virions in the same cell were observed in combinations of RMV-FG22, and TuMV-57, TuMV-TU2, or TuMV-R9 isolates haying virulence in radishes.

• 한국식물병리학회지
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• 제13권4호
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• pp.248-254
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• 1997

순무우 모자이크 바이러스 Ca 계통(TuMV-Ca)의 외피 단백질을 대장균 NM522 strain에서 발현시켰다. 발현된 바이러스 단백질은 한천젤 이중확산법, ELISA와 Western blotting을 이용하여 확인하였다. 외피단백질 발현 벡터(pGEX-Tu)의 구축은 IPTG induction site를 지니는 pGEX-KG에 TuMV-Ca 외피단백질 유전자를 결합하였다. 최적 단백질 발현 조건은 pGEX-Tu를 지니는 대장균을 액체 배지 1 ml당 $A_{595}$=0.1/ml의 농도로 접종한 후 2시간 뒤에 IPTG를 최종 농도를 1 mM로 조절하여 induction 시키는 경우였다. 합성된 목적 단백질은 발현 벡터의 특성상 GST (Glutathion S-Transferase) 단백질과 결합된 형태로 약 59 kDa의 단백질이었다. (uMV CP 33 kDa + GST 26 kDa.)

• Journal of Plant Biology
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• 제35권3호
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• pp.237-245
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• 1992

감자 (Solanum tuberosum L. cv. Superior)에서 cauliflower mosaic virus (CaMV) 35S promoter의 발현 양상 및 외래 유전자의 발현에 미치는 intron fragment의 효과를 조사하기 위하여 옥수수의 alcohol dehydrogenase 1-S (Adh1-S) intron 1의 249 base pairs 와 ${\beta}-glucuronidase$ (GUS) 유전자를 결합한, CaMV 35S/deleted Adh1 intron-GUS 구조의 유전자 전달벡터를 제조하고 이를 Agrobacterium tumefaciens를 매개로 형질전환을 유도하였다. 유전자 전달벡터인 pLS201는 17.7 kilobase pairs로서 형질전환의 초기 선별에 용이한 kanamycin 저항성 유전자와 GUS 유전자를 갖는 구조로 제조되었다. 형질전환된 개체의 조직화학적 분석 결과 CaMV 35S promoter에 의한 GUS 유전자는 모든 기관에서 발현되었고, 줄기 및 뿌리에서는 세포분열이 활발한 유관속 형성층을 중심으로 강한 발현을 나타내었다. GUS 유전자의 발현에 미치는 intron fragment의 효과를 조사하기 위하여 CaMV 35S/GUS 구조의 plasmid (pBI121)를 형질전환된 개체를 대조구하여 GUS 활성을 조사한 결과 pLS201의 잎, 줄기, 뿌리에서 각각 30, 34, 42배 높은 활성을 보여, 옥수수 탈수소 효소 유전자의 절단된 인트론이 GUS 유전자의 발현을 증가시킴을 알 수 있었다.

• 대한전자공학회논문지
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• 제22권6호
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• pp.28-33
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• 1985

집복회로제조기술을 이용하여 극소형이고 초경량이며 감도, 천택도 및 안정도가 뛰어난 ISFET(lon Sensitive Field Effect Transistor)를 제조하였다. 특히 사람의 혈액속에 나타나는 중요한 4가지 이온인 H+, K+, Na+, Ca++에 대하여 각가 선택적으로 반응하는 감지막을 형성하여 H+-ISFET, K+-ISFET, Na-ISFET 및 Ca++-ISFET를 제작하였다. 이들 4가지 ISFET의 감도는 각각 52mv/pH 42mv/pk, 59mv/pNa및 28mv/pCa로 나타났으며. 1초 미만의 빠른 응답을 보였다.

• Journal of Plant Biology
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• 제37권1호
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• pp.17-25
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• 1994

Two potato (Solanum tuberosum L.) cultivars were transformed by Agrobacterium tumefaciens harboring cauliflower mosaic virus (CaMV) 35S promoter and $\beta$-glucuronidase (GUS) gene. Expression patterns of the CaMV 35S promoter according to tissue types and developmental stages, and genetic stability of GUS gene were investigated in the clonal progenies of transgenic potatoes. Kanamycin-resistant shoot emerged from tuber disc after 4 weeks of culture, and root was induced 6 weeks after culture on the selection medium. Shooting frequency of cvs. Superior and Dejima were 43% and 27%, respectively. Mature transformants and their clonal progenies showed no phenotypical abnormality. GUS activity was expressed primarily at parenchymatous cells of phloem tissue around the vascular cambium in the stem and root, and higher activity was found at the apical meristem of shoot, root and adventious shoot bud. GUS activity was higher at tubers of young explants than at stored tubers. These facts indicate that expression level of the CaMV 35S promoter differed according to tissue types and developmental stages of the organs. The GUS gene was stably inherited to each clonal progeny and normally expressed.

• Journal of Plant Biotechnology
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• 제29권4호
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• pp.229-233
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• 2002

식물의 항산화력을 증진하여 식물병원균 저항성 작물을 개발하고자, 철분 결합 단백질인 대두의 ferritin 유전자를 CaMV 35S와 hsr203J promoter에 연결하여 감자에 형질전환하였다. PCR및 Northern분석에 의한 형질전환 감자에 ferritin 유전자가 존재하는 것과 이들 유전자 식물체내에서 발현되는 것을 확인하였다. ferritin 유전자를 담배 유래 병원균 특이 발현promoter인 hsr203J와 연결하여 획득된 형질전환 감자 식물체는 감자역병균 접종 후 24시간대에서 전사체 발현량이 가장 많았으며 그 후 줄어드는 경향을 나타내었다. 유전자 도입이 확인된 형질전환체 감자괴경의 철분 함량은 CaMV 35S와 ferritin 유전자 도입 형질전환체가 2.5배, hsr203J promoter와 ferritin 유전자 도입 형질전환체가 1.5배 각각 증가하는 것으로 나타냈다. 또한 이들 형질전환체는 감자 무름병균에 대한 저항성 증진효과를 관찰할 수 있었다.