• 식물조직배양학회지
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• 제25권4호
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• pp.225-229
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• 1998

상추의 종자 무균발아후 4일된 자엽조직을 GUS 유전자가 도입된 A. tumefaciens LBA 4404와 2일간 공동배양한 다음 0.1mg/L NAA, 1.0mg/L 2ip, 50mg/L kanamycin, 500mg/L carbenicillin이 첨가된 MS 배지에 배양하여 식물체를 재분화시켰다. PCR 분석결과 GUS 유전자가 형질전환된 식물체의 게놈상에 삽입되어 있음을 확인하였다. 해부학적 GUS 활성을 분석하여 형질전환된 식물체의 줄기, 잎 그리고 뿌리에서 GUS 유전자의 발현을 확인하였다. 형질전환체로 확인된 식물체를 자가수정시켜 얻어진 종자의 GUS 활성을 분석하여 GUS 유전자가 발현되는 것을 확인하였다.

• 식물조직배양학회지
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• 제25권1호
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• pp.37-43
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• 1998

꽃양배추 '은배' 종자를 무균 파종한 후 6일째된 하배축 조직을 BA 1㎎/L, sucrose 30㎎/L한천 8 g/L가 첨가된 MS 재분화배지에 1일간 전처리한 다음, PI promoter-GUS 융합 유전자가 도입된 Agrobacterium tumefaciens LBA 4404와 2일간 동일조성의 MS 액체배지에서 공동배양하여 carbenicillin 500 ㎎/L와 kanamycin 20 ㎎/L가 첨가된 MS 재분화배지로 옮겨 주었을 때 가장 많은 형질전환체를 얻을 수 있었다. PCR 분석결과, PI promoter-GUS 융합 유전자가 형질전환체의 게놈상에 삽입되어 있음을 확인하였다. Southern 분석결과, ECL-labelling된 PI promoter-GUS 융합 유전자 probe의 coding sequence와 동일한 것으로 판단되는 약 366bp 위치에서 밴드를 확인할 수 있었다. 그러나 형질 전환되지 않은 식물체에서는 이들 밴드를 확인할 수 없었다. 조직내 GUS 유전자의 활성은 잎부위에서부터 시작하여 엽병과 줄기의 관다발을 중심으로 나타났으며 상처의 정도가 심할수록 높은 편이었고 그 범위도 넓었다.

• Journal of Plant Biotechnology
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• 제31권4호
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• pp.279-283
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• 2004

백합 (Lilium longiflorum cv. Georgia) 화분의 생장과 agro-infiltration에 의한 일시발현에 대한 물리적, 화학적, 생물적 요인의 영향을 분석하였다 화분을 배지에 섞기 위한 물리적 과정이나 agro-infiltration을 위한 진공작업과정은 정상적 화분생장을 위하여 최소화되는 것이 바람직한 것으로 나타났다. 비교적 넓은 범위에서의 온도 (19 to 27$^{\circ}C$)나 pH(5.0 to 8.0)에서 화분의 생장이 유사하게 진행되었으며 화학적 요인으로서의 cefotaxime (300mg/L), acetosyringone (800 $\mu$M), syringealdehyde (800 $\mu$M) 등의 처리는 화분의 생장에 영향을 나타내지 않았다. 그러나 kanamycin의 경우 매우 심한 생장저해현상을 보였는데 25mg/L의 농도에서도 저해현상을 보이는 경우도 있었다. GUS유전자의 화분발현시 acetosyringone(200-400$\mu$M)의 처리에 의하여 그 효율이 약간 향상되는 것으로 나타났으나 syringealdehyde의 경우에는 효과가 없었다. 짧은 시간 내의 agro-infiltration과정과 이어서 18 hr의 화분 및 박테리아의 동시배양으로서도 acetosyringone의 첨가에 상관없이 화분에서의 GUS 일시 발현결과를 얻을 수 있었다.

• Journal of Plant Biotechnology
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• 제7권4호
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• pp.225-231
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• 2005

Five day old cotyledon explants of Cucumber (Cucumis sativus L) cv Poinsett 76 were cocultivated with two Agrobacterium strains (EHA105 and LBA 4404) each carrying GUS as the reporter gene and npt-II as the selection marker gene in the T-DNA region of the vector. Transformed shoots were selected at 150 mg/L kanamycin. A two day cocultivation coupled with $20\;{\mu}M$ acetosyringone increased the frequency (8.2 and 15.4 shoots) of GUS expression in the shoots of transformed plant. Among the two Agrobacterium strains, EHA 105 performed better than LBA 4404 in bringing two-fold increase in transformation efficiency (14%) than LBA 4404 (7.4%). PCR analysis was done to confirm the integration of T-DNA into cucumber genome.

• Current Research on Agriculture and Life Sciences
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• 제31권2호
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• pp.83-87
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• 2013

AtSAGT1 encodes a salicylic acid (SA) glucosyltransferase enzyme that catalyzes the formation of SA glucoside and SA glucose ester. Here, the AtSAGT1 gene expression patterns were determined in AtSAGT1 promoter::GUS transgenic Arabidopsis plants. As a result, the factors regulating the induction of AtSAGT1 were identified as pathogen defense response, wound response, exogenous application of SA, and jasmonic acid treatment.

• Journal of Plant Biology
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• 제38권4호
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• pp.365-371
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• 1995

Binary vectors, pBI-ActR1, pBI-ActF1 and pBSH-ActR1, were constructed using pGA642, the replication origin of pTi12 and the rice actin promoter. The sizes of pBI-ActR1, pBI-ActF1 and pBSH-ActR1 were 12.9 kb, 13.2 kb and 11.95 kb, respectively. These vectors containing a rice actin promoter followed by a GUS structural gene could induce stronly the expression of GUS gene in transformed rice cells. Rice explants from 3-4 day old seedlings after germinatin were cocultured with A. tumefaceins harboring pBI-ActR1, pBI-ActF1 or pBSH-ActR1, and then GUS expression in the explants was assayed. Transformation of rice explants by these binary vectors was tissue-specific, such that the meristematic regions of shoot apex, root and hypocotyl were transformed by these binary vectors.

• 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.

• Journal of Plant Biotechnology
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• 제38권4호
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• pp.308-314
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• 2011

Efficient transformation and regeneration methods are a priority for successful application of genetic engineering to vegetative propagated plants such as grape. In this study, methods for Agrobacterium tumefaciens-mediated transformation and plant regeneration of grapevine (Vitis vinifera) were evaluated. Tamnara, Heukgoosul, Heukbosek, Rizamat were co-cultivated with Agrobacterium strains, LBA4404 containing the vector pBI121 carrying with CaMV 35S promoter, GUS gene as reporter gene and resistance to kanamycin as selective agent. Seven percent of the maximum regeneration frequency was obtained from co-cultivated with explants from Rizamat with LBA4404 strain on selection medium with kanamycin. The addition of acetosyringone, 200 ${\mu}m$ in virulence induction step was a key factor for successful GUS reporter gene expression in grapevine transformation. Transgenic plants showed resistance to kanamycin and the GUS positive response in leaf ($T_0$) stem ($T_0$) and petiole ($T_0$).

• 한국작물학회:학술대회논문집
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• 한국작물학회 2000년도 춘계 학술대회지
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• pp.422-423
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• 2000

• Molecules and Cells
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• 제27권1호
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• pp.75-81
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• 2009

The Arabidopsis gene AtLEC (At3g15356) gene encodes a putative 30-kDa protein with a legume lectin-like domain. Likely to classic legume lectin family of genes, AtLEC is expressed in rosette leaves, primary inflorescences, and roots, as observed in Northern blot analysis. The accumulation of AtLEC transcript is induced very rapidly, within 30 min, by chitin, a fungal wall-derived oligosaccharide elictor of the plant defense response. Transgenic Arabidopsis carrying an AtLEC promoter-driven ${\beta}$-glucuronidase (GUS) construct exhibited GUS activity in the leaf veins, secondary inflorescences, carpel heads, and silique receptacles, in which no expression could be seen in Northern blot analysis. This observation suggests that AtLEC expression is induced transiently and locally during developmental processes in the absence of an external signal such as chitin. In addition, mechanically wounded sites showed strong GUS activity, indicating that the AtLEC promoter responds to jasmonate. Indeed, methyl jasmonate and ethylene exposure induced AtLEC expression within 3-6 h. Thus, the gene appears to play a role in the jasmonate-/ethylene-responsive, in addition to the chitin-elicited, defense responses. However, chitin-induced AtLEC expression was also observed in jasmonate-insensitive (coi1) and ethylene-insensitive (etr1-1) Arabidopsis mutants. Thus, it appears that chitin promotes AtLEC expression via a jasmonate- and/or ethylene-independent pathway.