[A quick method to estimate the T-DNA copy number in transgenic rice using inverse PCR (IPCR)].
In order to obtain single T-DNA copy transgenic rice, we have established a quick method to estimate the T-DNA copy number in transgenic rice using inverse PCR (IPCR). IPCR was used to amplify junction fragments, i.e. plant genomic DNA sequences flanking the known T-DNA sequences, which will help to estimate the T-DNA copy number in transgenic rice. We have analyzed 20 transgenic plants of 15 transgenic lines. Most plants (12) contain one integrated T-DNA copy per genome, 3 plants contain two and 1 plant contains 3 copies. In 4 transgenic plants no T-DNA copies could be detected using this method. The IPCR results were further tested by Southern analysis and sequence analysis.
Zhang
B
B
Shanghai Institute of Plant Physiology, Chinese Academy of Sciences, Shanghai 200032.
Huang
J Q
JQ
Wei
Z M
ZM
D004269:DNA, Bacterial / Q000032:analysis*; D018628:Gene Dosage; D012275:Oryza / Q000235:genetics*; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction / Q000379:methods*
D004269:DNA, Bacterial; C036483:T-DNA