| 摘要 |
NEW MATERIAL:A DNA coding rol A gene product derived from Ri plasmid held by Agrobacterium rhizogenes IFO 14555 strain, consisting of an amino acid sequence of the formula. USE:Plant breeding using recombinant DNA technique. PREPARATION:For example, Agrobacterium rhizogenes IFO 14555 strain is incubated in a medium followed by bacterium collection. The resulting bacteria is suspended in a buffer solution followed by addition of lysozyme and then making a phenol treatment to effect bacteriolysis followed by centrifugation, and the resulting supernatant is separated and spiked with ethanol, thus recovering DNA. The DNA is then treated with RNase followed by separating a plasmid fraction using cesium chloride-ethidium bromide density gradient centrifugation. Thence, this fraction is put to restriction enzyme treatment and cloning using a probe through colony hybridization, thus obtaining the objective DNA coding rol A gene product, consisting of an amino acid sequence of the formula. |
