• Journal of Plant Biotechnology
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• 제35권1호
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• pp.41-45
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• 2008

Laccase (EC 1.10.3.2) is a small group of enzymes that catalyze the oxidation of a broad range of phenolic compounds including hazardous and recalcitrant pollutants in the environment. This study attempted to develop an efficient system for production of a recombinant laccase by chloroplast genetic transformation of tobacco. Chloroplast transformation vector was constructed and introduced into the tobacco chloroplast genome using particle bombardment. Chloroplast-transformed plants were subsequently regenerated. PCR and southern blot analyses confirmed stable integration of the laccase gene into the chloroplast genome. Northern blot analysis revealed that mRNA of the laccase gene was highly expressed in chloroplast-transformed plants.

• Journal of Plant Biotechnology
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• 제30권2호
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• pp.143-149
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• 2003

This experiment was carried out to confirm the stability of bar gene introduced into petunia plant through Agrobacerium-mediated transformation. Twenty-five transgenic plants T$_{0}$ plants, back cross (BC$_1$) populations to wild type and F$_1$plants between different T$_{0}$ plants were prepared, and polymerase chain reaction(PCR), PCR-Southern blot analysis, and field test with 0.1% Basta treatment were done. The results of PCR, PCR-Southern blot hybridization, and field test indicated that NPTII and bar gene introduced into the genome of petuina plants were stably transmitted to their progenies, and conferred the plants resistance to herbicide, Basta.sta.

• Journal of Plant Biotechnology
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• 제34권3호
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• pp.271-275
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• 2007

엽록체 형질전환된 식물체를 얻으려면 먼저 세포수준에서 모든 엽록체가 형질전환되어야 하는데, 세포내에는 많은 수의 엽록체가 존재하므로, 엽록체 형질전환 벡터가 전이되어 형질전환된 엽록체는 선발배지에서 선택적으로 분열을 계속하고 형질전환되지 않은 엽록체들은 분열을 하지 못하게 되어, 결국 해당 세포내의 모든 엽록체가 형질전환된 상태에 이르게 된다. 따라서 만일 해당 세포내에 엽록체의 수가 적으면 그만큼 효율적으로 엽록체 형질전환을 할 수 있을 것이다. 본 연구에서는 담배의 FtsZ 유전자를 핵형질전환법으로 과잉 발현시킴으로써 엽록체의 분열이 저해되어 엽육세포내에 거대한 엽록체 3-5개를 가진 담배식물체의 엽육조직을 이용하여, 엽록체 형질전환을 한 결과, 엽록체 형질전환 빈도가 약 40% 증가되었다.

• Journal of Plant Biology
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• 제38권2호
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• pp.203-209
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• 1995

담배(Nicotiana tobacum L. cv. Wisconsine 38) 잎 절편을 해녀콩(Canavalia lineata)의 leghemoglobin(Lb) cDNA를 포함하는 Agrobacterium과 함께 배양한 후, 0.5mg/L BAP, 0.1mg/L ${\alpha}-NAA$와 200mg/L kanamycin, 500 mg/L carbenicillin을 포함하는 MS 배지에서 선별하여 7개의 재분화 개체를 얻었다. 이로부터 분리한 게놈 DNA에 대한 Southern 혼성화 반응과 PCR 결과로 Lb cDNA가 담배의 게놈에 삽입되었음을 확인하였다. 형질전환된 담배로부터 분리한 RNA에 대한 northern 혼성환 실험 결과 약 1,000 nt의 RNA가 혼성화 반응을 보였으며, 총 RNA를 주형으로 합성한 1차 가닥의 cDNA를 PCR로 증폭한 결과, Lb cDNA와 혼성화 반응을 보이는 0.5 kb의 DNA가 증폭되었다. 콩의 Lb에 대한 다군항체(polyclonal antibody)를 사용하여 단백질 면역 항체 반응을 실시한 결과, Lb로 판단되는 약 15.8 kD의 위치에서 혼성화 반응이 나타났다. 이상의 결과로 해녀콩 Lb cDNA가 형질전환된 담배의 게놈에 삽입되었을 뿐만 아니라 mRNA로 전사되어 Lb 단백질로 해독되었음을 알 수 있었다.

• 식물병연구
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• 제19권3호
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• pp.177-182
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• 2013

Resvestrol has been known to inhibit bacterial and fungal growth in vitro, and can be accumulated in plant to concentrations necessary to inhibit microbial pathogens. Hence, stilbene synthase gene has been used to transform to synthesize resveratrol in heterologous plant species to enhance resistance against pathogens. In the present study, we investigated the antimicrobial activities of resveratrol and piceid to bacterial and fungal pathogens, which causing severe damages to rice plants. In addition, disease resistance was compared between transgenic rice varieties, Iksan 515 and Iksan 526 transformed with stlibene synthase gene and non-transgenic rice varieties, Dongjin and Nampyeong. Minimum inhibitory concentration of resveratrol for Burkolderia glumae was 437.5 ${\mu}M$, and the mycelial growth of Biplaris oryzae was slightly inhibited at concentration of 10 ${\mu}M$. However, other bacterial and fungal pathogens are not inhibited by resveratrol and piceid. The expression of the stilbene synthase gene in Iksan 515 and Iksan 526 did not significantly enhanced resistance against bacterial grain rot, bacterial leaf blight, sheath blight, and leaf blight. This study is the first report on the effect of resveratrol and piceid against pathogens of rice plant, and changes of disease resistance of transgenic rice plants transformed with stilbene synthase gene.

• Journal of Plant Biology
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• 제35권3호
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• pp.229-235
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• 1992

담배 식물체에서 Arabidopsis thaliana에 존재하는 3가지 다른 엽록소 a/b 결합 단백질 (cab) 유전자 promoter들의 발현을 조사하였다. 이들 promoter의 활성은 이들 promoter에 결합된 reporter gene의 식물인 CAT (chloramphenicol acetyltransfer-ase)의 활성으로 측정하였다. 담배엽에서 cab promoter의 활성은 cab1, cab2, cab3의 순이었고, 이들 잎에서 유도한 callus나 shoot에서도 동일한 양상을 보였다. 이들 3가지 promoter의 발현은 상부엽에서 하부엽보다 높은 활성은 보였으며 개체간의 높은 변이폭은 뿌리내리기를 통하여 무성증식된 식물체를 사용하여 줄일 수 있었다. 이들은 또한 기관 특이성을 보여 줄기 보다 잎에서의 활성이 높았고 뿌리에서는 거의 발현되지 않았다. 그러나 cab1 promoter의 경우, 다른 두 cab promoter들과는 다르게, 잎에 대한 줄기에서의 상대적인 활성이 높았으며, CAT 활성을 단위 단백질당을 표시하는 대신 단위 엽록소당 활성으로 나타내었을 때 줄기와 잎에서 활성의 차이가 관찰되지 않았다. 즉 cab2와 cab3는 광합성 기관 특이성을 보이는 반면 cab1의 경우는 이러한 특이성을 나타내지 않았다. 이러한 현상은 잎에서 유도한 shoot가 줄기와 잎으로 분화되는 과정에서도 관찰되었다.

• 한국초지조사료학회지
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• 제17권4호
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• pp.379-386
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• 1997

대만산 배추 (Brassica campestris M.)로부터 분리한 BcHSPI7.6 cDNA(내열성 유전자)를 pBKSl-l vector에 subcloning하므로서, NPTII 유전자와 P35S-HSPI7.6 cDNA를 가지는 pBKH4 재조합 플라스미드를 제작하였다. 이들 플라스미드를 갖는 A. tumefaciens LBA4404로서 담배잎 단편을 24시간 동안 공배양하므로서 감염시켰으며, 이들 유전자로 형질전환된 shoot는 $100\;{\mu\textrm{g}}/ml$의 가나마이신을 첨가한 MS-n/B 배지에서 선발하였다. 담배((Nicotiana. tabacum)의 열에 대한 치사온도는 $50^{\circ}$에서 15분 이상이었으며, BcHSPI7.6 cDNA를 갖는 형질전환된 식물체는 이 온도에서 내열성을 나타내었다. 식울체의 형질전환 여부는 ${\alpha}^{_32}P$로 표지한 BcHSPI7.6 cDNA 단편을 probe로 이용해서 Southern blot hybridization을 실시하므로서 확인하였다. BcHSPI7.6 cDNA의 발현정도는 Northern blot 분석과 이중면역확산 방법으로 확인하였다. 본 연구에서, 담배에 도입한 BcHSPI7.6 cDNA는 내열성과 관련이 있는 유전자로서, HSPI7.6 단백질은 식물체를 열에 의한 손상으로부터 방지해 주는 protector 역할을 하는 것 으로 사료된다.

• Journal of Biosystems Engineering
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• 제23권5호
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• pp.491-498
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• 1998

Transplanting process during the tissue culture of potato seedlings is costly, since the cost of highly skilled labor working in the sanitary environment takes up about 60-70% of the production cost. The objective of this study was to develop a soft gripper of a transplanting robot system for the labor-saving tissue culture. The prototype of the soft gripper was consisted of power-transmitting part finger and plant contacts. The power transmitting part transformed the rotating motion of a step motor to the reciprocating motion of the finger. Plant stems used in the test were potato seedlings cultured for six weeks. The dimensional characteristics of cultured seedlings, the compressive strengths of the stems, the extractive force from the culture medium and the gripping force of the finger were measured. A proper gripping force was found to be 0.343N at the extractive force of 0.41N when the plant contacts were made of silicon. Sixteen plants out of 70 trials were tangled with others, resulting in the success rate of 77.1%.

• 한국연초학회지
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• 제21권2호
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• pp.152-159
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• 1999

A CAB cDNA clone(pKGCAB) encoding the light harvesting chlorophyll a/b binding protein of the semi-shade plant, Korean ginseng(Panax ginseng C. A. Meyer) was isolated by the one-way path random sequencing of ginseng cDNA library clones and transgenic tobacco plants(Nicotiana tabacum NC82) were produced by the transformation of this ginseng CAB gene in use of Agrobacterium tumefaciens LBA4404. The CAB gene showed type 1 structure of LHCP-II, 84% similarity in nucleotide sequence and 92% in amino acid sequence to that of Nicotiana tabacum CAB40, respectively. Seed germination and initial growth of the transgenic tobacco plants transformed with the cDNA fragment were accelerated under low light intensity compared with those of normal tobacco plant, that may result from the higher light sensitivity of the transgenic plants than that of the normal.

• 한국연초학회지
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• 제20권1호
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• pp.57-65
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• 1998

This study was conducted to develop a resistant tobarro against Potato virus Y (PVY) by transformation of the plants with genetically engineered viral genes. The complimentary DNAs (cDNAS) of potato virus Y-necrosis strain (PVY-Vn) replicase mutant genes (3'-deleted, 5'-deleted and ADD-mutant Nlbs) were synthesized through RT-PCR by using purified PVY-VN RNA and synthesized primers, and cloned in the sense orientation into a plant expression vector (pMBPI), The cDNAS of the genes were transferred into Agrobacterium tumefaciens LBA 4404, and then transformed into tobacco (Nicotiana tabacum cv. Burley 21) plants. Regenerated plants were tested for PVY resistance by inoculation test; 13 transgenic plants including 7 for 3'-deleted Nlb, 3 for 5'-deleted Nlb, and 3 for ADD-mutant Nlb appeared to be resistant at 4 weeks after inoculation with PVY-VN. Among the 13 transgenic tobacco plants, 8 plants had no symptom up to 14 weeks after inoculation. The progenies ($T_1$) from self-fertilization of the transgenic lines varied 0.0% to 81.2% in their resistance (% of resistant plants). The analysis of Nlb-31deleted, -5'deleted and -ADD mutant in the $T_1$ plants by polymerase chain reaction (PCR) showed that Nlb-3'deleted, -5'deleted and -ADD mutants were detected in all of the resistant plants. These results suggest that the PVY resistance was inherited in the $T_1$ generation.