Detection of rol Genes of Ri Plasmids by PCR Method and Its Application to Confirmation of Transformation.

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  • ＰＣＲ法によるＲｉプラスミドｒｏｌ遺伝子の検出と形質転換確認への応用
  • PCRホウ ニ ヨル Ri プラスミド rol イデンシ ノ ケンシュツ ト

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Four oligonucleotide primers which showed the same homologous sequences in rol genes between mikimopine and agropine type Ri plasmids were synthesized. When two pairs of the primers were used for polymerase chain reaction (PCR) with the total DNA of ten strains of A. rhizogenes as templates, fragments with a same length were amplified in nine strains.<br>Transgenic tobacco plants were obtained from hairy roots transformed with two strains of A. rhizogenes, A4 and NIAES1724. Amplification of a single fragment originated from inserted DNA into plant genome occured by the same reactions. That these results are not due to contamination by bacterial DNA was confirmed by using PCR with border sequences as primers.

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