Korean Journal of Plant Tissue Culture (식물조직배양학회지)

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Advanced Regeneration and Genetic Transformation of Lycium chinense Harboring Salt Tolerance Genes

구기자나무 (Lycium chinense)의 효과적인 재분화 및 내염성 유전자가 도입된 형질전환체의 개발

  • Published : 2001.01.01
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Abstract

Bet A and Bet B genes related to salt resistance were introduced into Lycium chinense through high efficiencies of plant regeneration. The explants were precultured on the shooting medium which is consisted of MS medium added 1mg/L kinetin and 0.05mg/L IBA for 2 days. After pre-culture, they were immersed in LB media containing Agrobacteria tumefaciens harboring Bet genes, and cultured on the same medium. Putative transformants could be selected after cocultivation of the explants with Agrobacteria on the shooting medium supplemented with 30mg/L kanamycin. The presence of both Bet A and Bet B genes from the transgenic plants were confirmed by PCR amplification with the gene specific primers and subsequent PCR-Southern blot with labeled Bet genes probe. The expression of Bet A and Bet B genes in the transgenic plants were observed by RT-PCR method.

구기자나무의 효과적인 재분화조건을 바탕으로 염류내성유 전자인 Bet A와 Bet B유전자의 도입을 시도하였다. 구기자나무의 절편체를 재료로 kinetin 1 mg/L, IBA 0.05 mg/L가 첨가된 MS배지에 2일간 전배양한 후 Agrobacterium과 공조배양 및 선발배지에서의 배양으로 kanamycin에 내성을 갖는 잠정적인 형질전환체를 유도하였다. 형질전환체는 PCR 기법 및 Southern blot분석으로 Bet A와 Bet B 유전자 전이를 확인하였고, 도입된 유전자의 발현은 RT-PCR 방법을 사용하여 확인하였다.

Keywords

References

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