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Yi Chuan ; 40(8): 676-682, 2018 Aug 16.
Article in Chinese | MEDLINE | ID: mdl-30117423

ABSTRACT

The availability of T-DNA insertion sites is very important for plant functional genomics research and the screening and identification of transgenic plants. However, the present protocols for identifying T-DNA insertion sites, like reverse PCR and semi-random primer PCR, are not only complex and time-consuming, but also inefficient. In this paper, a DNA pool of three transgenic plants was sequenced by whole-genome resequencing, and four T-DNA insertion sites were identified by blasting using transgenic T-DNA sequences. After PCR and Southern blot analysis, the T-DNA insertion sites of the three transgenic plants were successfully confirmed, and one of the transgenic plants showed two insertion sites. In conclusion, this study established a simple, reliable and efficient method for obtaining T-DNA insertion sites in transgenic plants.


Subject(s)
DNA, Bacterial/genetics , Mutagenesis, Insertional , Oryza/genetics , Plants, Genetically Modified/genetics , Base Sequence , DNA Primers/genetics , DNA, Plant/genetics , Genome, Plant , Polymerase Chain Reaction , Sequence Analysis, DNA
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