Korean Journal of Plant Tissue Culture (식물조직배양학회지)

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Transformation of PAT gene into Lettuce (Lactuca sativa L.) using Agrobacterium tumefaciens

Agrobacterium tumefaciens를 이용한 상추 (Lactuca sativa L.)의 PAT유전자 형질전환

  • Published : 2001.07.01
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Abstract

Agrobacterium tumefaciens MP90 harboring PAT (phosphinothricin acetyltransferase) and NPTII-GUS gene were used for the genetic transformation of lettuce (Lactuca Sativa L.). Shoot regeneration from cotyledon explants were obtained from the MS medium supplemented with 0.1 mg.L$^{-1}$ NAA, 1.0 mg.L$^{-1}$ 2ip, 50 mg.L$^{-1}$ kanamycin and 500 mg.L$^{-1}$ carbenicillin after cocultivation with A. tumefaciens for 2 days. Kanamycin resistance test of transgenic plants indicated that the NPTII gene was integrated into the lettuce genome and was stably expressed. PCR and northern blot analysis indicated that bialaphos resistance gene (PAT) was stably integrated into the lettuce genome. The transgenic plant sprayed with Basta (1500x) remained healthy with continuous growth, while the control group exhibited fatality.

상추에 제초제 저항성을 도입하기 위해 상추 자엽 조직을 NPTII-GUS 및 제초제 저항성 유전자 (PAT)가 삽입된 A. tumefaciens MP 90과 2일간 공동배양한 다음 0.1 mg.L$^{-1}$ NAA, 1.0 mg.L$^{-1}$ 2ip, 50 mg.L$^{-1}$ kanamycin, 500 mg.L$^{-1}$ carbenicillin을 첨가한 MS배지에 배양하여 식물체를 재분화시켰다. 재분화 식물체는 kanamycin 내성 검정을 통해 NPTII 유전자의 도입과, PCR, Northern blot 분석을 통해 제초제 저항성 유전자가 식물체의 게놈상에 삽입된 형질전환체임이 확인되었다. 또한 1500배액의 농도로 Basta를 살포한 결과 살포 10일 후 대조 식물체는 완전히 고사하는 데 반해 형질전환체는 지속적인 생육을 보여 얻어진 형질전환 상추가 제초제 저항성 개체임이 확인되었다.

Keywords

References

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