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RISS 인기검색어
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- 저자 : 이효연 (검색결과 174 건)
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Expression of Human Lactoferrin Gene in Transgenic Rice(Oryza sativa L.)
이효연,Yong-Eok Lee,Seong-Eun Oh,Satoshi Nishiguchi,Key-Zung Riu,송인자,박신영,Jin-Hyoung Lee,Il-Gi Kim,서석철,Seong-Lyul Rhim,임평옥 한국식물생명공학회 2007 JOURNAL OF PLANT BIOTECHNOLOGY Vol.34 No.2
Lactoferrin is an 80-kDa iron-binding glycoprotein known to exert many biological activities, such as facilitating iron absorption and having antimicrobial and anti-inflammatory effects. Rice can be a useful target for edible food plants to introduce human lactoferrin, because it has lower allergenicity and is likely to be safer than microorganisms or transgenic animals. A cDNA fragment encoding human lactoferrin (HLF) driven by the maize polyubiquitin promoter, along with herbicide resistance gene (bar) driven by CaMV 35S promoter, was introduced into rice (Oryza sativa L. cv. Dong Jin) using the Agrobacterium -mediated transformation system. Putative transformants were initially selected on the medium containing bialaphos. The stable integration of the bar and HLF genes into transgenic rice plants was further confirmed through polymerase chain reaction (PCR) and Southern blot analyses. The expression of the full length HLF protein from various tissues such as grains and young leaves of transgenic rice was verified by Western blot analysis. Analysis of progeny also demonstrated that introduced genes were stably inherited to the next generation at the Mendelian fashion.
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李孝淵 順天大學校 農業科學硏究所 1994 農業科學硏究 Vol.- No.-
The herbicide bialaphos is a potent inhibitor of glutamine synthetase in higher plants. A bialaphos resistance (bar) gene encoding for an acetyltransferase was isolated from genomic DNA of Pseudmonas syringae pv tabaci. The bar gene was logated to the binary vector pBO121. Postils of tobacco plants were treated with the bar gene containing plasmid DNA at various times after pollination. When the treatment was applied at 30 and 40 h after pollination, a number of transgenic plants wrer obtained. Primary transforments (T_0 generation)and their progenies (T_2, T_3) were resistant to both bialaphos and kanamycin at a dosage lathal to untransformed control plants. Stable integration of bar gene into chromosomal DNA was proven by Southern blot analysis of genomic DNA isolated from T_1 progenies. These results show that the bialaphos resistant plants could be obtained by treatment to postils with the exogenous bar gene through the fertilization cycle of tobacco. For the gene taffing of Armoracia rusticana, maize contrilling element Ac and Ds were introduced into A. rusticana via Agrobacterium-mediated transformation method. We established an efficient in vitro regeneration and transformation system for gene transfer in A. rusticana. The optimum in vitro regenrration condition has been obtained from leaf, petiole and root organs on modified MS medium supplemented with NAA 0.1㎎/ℓ plus BA 1.0㎎/ℓ for direct shooting and with free growth regulators for root induction. For transformation, the leaf, petiole and root explants of A. rusticana were cocultivated with Agrobacterium tumefaciens, LBA4404 which carries a bianry vector pEND4K containing maixe contrilling element Ac or Ds, respectively. Selections were performed in the shoot induction medium supplemented with 100㎎/ℓ kanamycin, and 500㎎/ℓ carbenicillin. Transformation frequency showed about 8∼10% in case of leaf disks. PCR and Southern blot analyses showed that the Ac and the Ds elements wee integrated into the chromosome of donor plants.
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EMS처리에 의한 한란의 엽록소 결핍 돌연변이 식물체의 유도
이효연,정재성,이종석 한국식물생명공학회 1998 식물생명공학회지 Vol.25 No.3
Chlorophyll mutants were produced by treating the rhizome of Cymbidium kanran with mutagen, EMS(ethyl methan sulfonate). The germination ratio of Cymbidium kanran seeds was 5.5 times higher when the seeds were treated with ultrasonic treatment for 20 minutes than untreated control. Fifty to sixty percent of the rhizomes became dark brown when they were cultured in a liquid growth medium containing 0.2% EMS for three weeks. When the dark-brown rhizomes were cultured in a solified MS medium, new rhizomes were developed from a part of the old ones. Chlorophyll mutant rhizomes were obtained from a meristem tissue by a subculturing the cuts of these new rhizomes for a year. Of the chlorophyll mutants, a zigzag-striped type of rhizome was dominant and light-yellow and albino ones were also produced. While the zigzag-striped type rhizomes were differentiated into green and striped plant, the light yellow and the white rhizomes produced yellow-striped and albino plants repectively.These results indicate that the EMS treatment on the rhizome is an effective means to induce a chlorophyll mutant. We believe that this method may be useful to produce variegated plants chlorophyll mutants from other orchids. 한란(Cymbidium kanran)의 근경에 돌연변이 물질인 EMS를 처리하여 엽록소변이 식물체를 유도하였다. 한란종자를 발아시켜서 근경을 유도할 때 종자에 40분간 초음파 처리한 것이 무처리구에 비하여 발아율이 5.5배 정도 높았다. EMS 0.2%를 포함한 근경증식용 액체배지에서 3주간 배양하였을 경우에 50-60%정도의 근경이 갈변되었다. 갈변된 근경을 근경증식용 고체배지에서 배양하였을 경우에 근경조직의 일부분으로부터 새로운 근경의 증식의 관찰되었고, 이러한 근경조직을 절단하여 1년간 계단배양하던중에 분열조직의 일부분에서 색소변이 근경이 관찰되었다. 색소변이 근경은 나선형의 줄무늬가 가장 많았고, 그 외 엷은 노란색과 흰색의 근경이 분리되었다. 나선형의 근경에서 유도된 식물체는 잎에 무늬가 전혀 없는 녹색의 식물체 또는 줄무늬가 들어 있는 식물체가 유도되었다. 그러나 엷은 노란색 근경에서 유도된 식물체는 노란색 줄무늬가 있는 식물체로, 흰색 근경으로부터 유도된 식물체는 백화된 식물체로만 분화하였다. 이상의 결과 한란근경을 이용한 색소변이식물체의 유도에 있어서 배양중인 근경에 EMS처리가 효과적인 것을 보여주었다. 이러한 방법을 다른 식물에 적용하면 색소변이 식물체를 개발할 수 있으리라 생각된다.
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Agrobacterium tumefaciens를 이용한 bialaphos 저항성 형질전환 벼의 개발
이효연,이춘환,김호일,한원동,최지은,김진호,임용표 한국식물생명공학회 1998 식물생명공학회지 Vol.25 No.4
비선택성 제초제인 bialaphos는 고등식물에 있어서 glutamine 합성을 억제 하여 식물체를 고사시킨다. Acetyltransferase에 의해 encoding된 bialaphos저항성 유전자는 세균 Streptomyces hygroscopicus SF1239로부터 cloning된것을 사용하였다. Bialaphose 저항성 유전자를 Agrobacterium 감염법을 이용하여 국내에서 재배되는 벼(동진)에 도입한 결과 약30%정도의 형질전환 식물체를 얻을 수 있었다. $\textrm{T}_{1}$ 세대의 17개체는 hygromycin과 bialaphos에 대한 저항성 유전형질이 3:1로 분리되었다. 또한 Southern 분석을 실시한 결과 wild type의 식물체에서는 Bar 유전자의 검출을 볼 수 없었으나 형질전환 식물체의 경우 Bar 유전자의 검출이 가능하였다. $\textrm{T}_{3}$세대의 형질전찬 식물체와 wild type의 식물체를 포장상태에서 비선택성 제초제인 바스타를 살포하고 3주 후에 관찰한 결과 형질전환 식물체는 외형적으로 아무런 피해를 받지 않고 정상적으로 생장하였으나, wild type의 식물체와 잡초는 모두 고사하였다. 이상의 결과를 종합해보면 hygromycin과 bialaphos 저항성 유전자는 Agrobacterium감염법을 이용하여 단자엽 식물인 벼에 도입할 수 있다는 것을 보여준 것이며, 또한 bialaphos 저항성 유전자가 식물에 도입됨으로써 비선택성 제초제에 대한 저항성 식물을 개발 할 수 있다는 것을 보여 주었다. The bialaphos is a potent inhibitor of glutamine synthease in higher plants and is used as a non-selective herbicide. We have used the bialaphos resistant gene(Bar) encoding for an acetyltransferase isolated from Streptomyces hygroscopicus SF1293. Callus derived from mature seeds of rice(Oryza sativa L. cv. Dong Jin) were co-cultivated with Agrobacterium tumefaciens EHA101 carring a plasmid pGPTV-HB containing genes for hygromycin resistance (HygR) and Bar. Transgenic plants showing in vitro resistance to 50 mg/L hygromycin and 10 mg/L bialaphos were obtained by using a two-step selection/regeneration procedure. Transformation efficiency of rice was about 30% which was as high as reported in other dicotyledons. Progenies ($\textrm{T}_{1}$ generation) derived from primary transformant of 17 lines were segregated with a 3 resistant : 1 sensitive ratio in medium containing hygromycin and bialaphos. Stable integration of Bar gene into chromosomal DNA was proven by Southern blot analysis of genomic DNA isolated from $\textrm{T}_{2}$ progenies. Transgenic plants ($\textrm{T}_{3}$) grown in the field were resistant to bialaphos (Basta) at a dosage lethal to wild type plants.
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