발표요지 : 벼의 5- Methyltryptophan 저항성주의 선발과 세포융합

이효연,김진호,김학진,귀곡수소 (龜谷壽昭) 한국식물생명공학회 ( 구 한국식물조직배양학회 ) 1993 한국식물생명공학회 춘계학술연구발표회 Vol.1993 No.-

RAPD 표지인자를 이용한 흑오미자의 자웅동주 및 자웅이주 식물의 동정

이효연,한효심,이갑연,한상섭,정재성 한국식물생명공학회 1998 식물생명공학회지 Vol.25 No.5

본 연구는 국내의 경우 제주도의 일부지역에서만 자생하는 흑오미자(Schisandra nigra)를 RAPD법을 이용하여 자웅동주 및 자웅이주 식물의 특이적 Marker를 탐색하고저 실시 하였다. 10-mer로 구성된 80종류의 random primer를 사용하여 흑오미자를 분석한 결과 기존의 재배되고 있는 오미자(Schisandra chinensis) 또는 남오미자(Kadsura japonica)와는 다른 band pattern을 보여 주었다. 흑오미자의 자웅동주. 암그루, 숫그루의 3품종을 상기와 동일하게 80개의 primer를 사용하여 RAPD를 분석한 결과, 5종류의 random primer(OPA-17, OPA-19, OPB-3, OPB-9, OPB-16)에 대해서는 각 품종에 대한 특이적인 band가 검출되었다. 숫그루, 암그루 식물과는 다르게 자웅동주 식물은 3개체(1호 2호, 3호)간에도 서로 다른 band pattern을 보이는 특징을 갖고있다. 숫그루 특이적인 band pattern은 OPB-3 primer을 이용할 경우 750bp에서 검출되었고, 암그루는 OPA-19 primer에서 950bp, 1690bp와 OPB-3 primer의 경우 700bp에서 자웅동주 및 숫그루에서 나타나지 않는 band가 검출되었다. 이러한 결과는 흑오미자의 숫그루 및 암그루에서 나타난 특이적인 band가 유묘시기에 암ㆍ수 개체를 조기에 구별하는 genetic marker로 사용할 수 있으리라 기대된다. RAPD (Random Amplified Polymorphic DNA) analysis was conducted to Schisandra nigra plants in order to select the specific markers for monoecious and dioecious individuals. RAPD results using eighty random 10-mer primers revealed that S. nigra had a different banding pattern from S. chinensis and Kadsura japonica. When DNA isolated from leaves of monoecious and dioecious plants were used as PCR template, only five primers, OPA-17, OPA-19, OPB-03, OPB-09 and OFB-16, showed polymorphic band patterns. No variation in banding profiles within male or female individuals was observed when these five primers were used whereas three monoecious plants (No 1, No 2 and No 3) showed different banding patterns one another, A 750 bp segment was amplified by primer OPB-3 from male individuals. On the other hand, two segments, 950 bp and 1690 bp, with OPA-19 and 700 bp of segment with OPB-3 were amplified in female individuals. These result indicate that the specific buds of male and female S. nigra could be used as genetic markers for the early discrimination of male and female individuals.

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.

길위에 선 사람들

이효연 열린전북 2002 열린전북 Vol.38 No.-

受粉·受精 시기를 이용한 형질전환 식물의 개발

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.

Ethylene Imine 처리에 의해서 얻어진 벼의 5-methyltryptophan 저항성주의 특성

이효연,김진호,김학진,권병선,구곡수소 한국작물학회 1992 한국작물학회 학술발표대회 논문집 Vol.1992 No.-

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.

Agrobacterium tumefaciens를 이용한 bialaphos 저항성 형질전환 벼의 개발

이효연,이춘환,김호일,한원동,최지은,김진호,임용표 충남대학교 생물공학연구소 1999 생물공학연구지 Vol.7 No.-

비선택성 제초제인 bialaphos는 고등식물에 있어서 glutamine 합성을 억제하여 식물체를 고사시킨다. Acetyltransferase에 의해 encoding된 bialaphos 저항성 유전자는 세균 Streptomyces hygroscopicus SF1239로부터 cloning된 것을 사용하였다. Bialaphose 저항성 유전자를 Agrobacterium 감염법을 이용하여 국내에서 재배되는 벼(동진)에 도입한 결과 약 30%정도의 형질전환 식물체를 얻을 수 있었다. T_1세대의 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. T_3 세대의 형질전환 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 Agrobacterium 감염법을 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 isloated from Streptomyces hygroscopicus SF1293. Callus derived from mature seeds of rice (Oryza sativa L. cv. Dong Jin) were co-cultivated with Agrobacterium tumefacieus 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 (T_1 generation) derived from primary transformant of 17 lines were segreagated 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 T_2 progenies. Transgenic plants (T_3) grown in the field were resistant to bialaphos (Basta) at a dosage lethal to wild type plants.

[행복 일기] 요리로 지지고 볶는 수학공부

이효연 샘터사 2009 월간 샘터 Vol.473 No.-

Direct transformation 을 이용한 제초제 저항성 유전자의 도입

이효연 한국생약학회 1993 생약학회지 Vol.24 No.1