[Spotting the positions of T-DNA on the genome for Magnaporthe grisea transformants and study on the mode of T-DNA insertion].
Using thermal asymmetric interlaced PCR (TAIL-PCR), 39 specifical fragments of Magnaporthe grisea genomic DNA flanked on the T-DNA were successfully amplified from 37 M. grisea transformants randomly selected from the mutants induced by T-DNA insertion in our laboratory. These fragments were sequenced and then compared by BLAST with the sequences of M. grisea genomic DNA published in netwwork. T-DNA insertion positions for 17 transformants were spotted on the genome of M. grisea. Of all the 39 amplified specifical fragments, 19 were M. grisea genomic DNA and 20 contained the vector backbone sequences. Of the 19 M. grisea genomic DNA fragments, 10 flanked on the right and 9 on the left border of T-DNA inserted. Of the 10 M. grisea genomic DNA fragments flanked on the right border of T-DNA, 9 had a 102bp identical sequence. However, the 7 fragments flanked on the left border of T-DNA did not have this regularity. The above results demonstrated that T-DNA right nick positions were relatively fixed on the M. grisea genomic DNA, similar to the ones on plant genomic DNA. The spotted positions of T-DNA on the genome for 17 M. grisea transformants established a solid foundation for further functional genomic research of M. grisea.
Lin
Chun-hua
CH
Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou 571737, China. lin3286320@126.com
He
Chun-ping
CP
Wang
Kui-di
KD
Liao
Qi-heng
QH
Zheng
Fu-cong
FC
D004269:DNA, Bacterial / Q000235:genetics*; D016681:Genome, Fungal; D020082:Magnaporthe / Q000235:genetics*; D016254:Mutagenesis, Insertional; D014170:Transformation, Genetic
D004269:DNA, Bacterial; C036483:T-DNA