Korean Journal of Plant Tissue Culture (식물조직배양학회지)

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Expression of $\beta$-Glucuronidase (GUS) Gene in Transgenic Lettuce (Lactuca sativa L.) and Its Progeny Analysis

형질전환된 상추내에서 GUS 유전자의 발현 및 후대검정

  • Published : 1998.07.01
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Abstract

Agrobacterium tumefaciens LBA 4404 harboring binary vector pBI 121 was used for genetic transformation of lettuce(Lactuca Sativa L.). Optimal shoot regeneration from cotyledon explants was obtained in MS medium supplemented with 0.1mg/L NAA and 1.0 mg/L 2ip. In this condition, cotyledon explants were cocultivated with A, tumefaciens for 2 days, and then transferred to selection medium supplemented with 50 mg/L kanamycin and 500 mg/L carbenicillin. These explants were subsequently subcultured every 2 weeks on shoot induction medium. PCR analysis indicated that the GUS gene was stably integrated into the nuclear genome of lettuce. Histochemical analysis based on the enzymatic activity of the CUS protein showed that GUS activity was associated with vascular tissue in leaves and roots. Progenies of Ro plants demonstrated a linked monogenic segregation for GUS gene.

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.

Keywords

References

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