Yeast-based recombineering of DNA fragments into plant transformation vectors by one-step transformation.
T-DNA binary vectors are often used in plant transformation experiments. Because they are usually very large and have few restriction sites suitable for DNA ligation reactions, cloning DNA fragments into these vectors is difficult. We provide herein an alternative to cloning DNA fragments into very large vectors. Our yeast-based recombineering method enables DNA fragments to be cloned into certain types of T-DNA binary vectors by one-step transformation without the requirement of specific recombination sites or precisely positioned restriction ends, thus making the cloning process more flexible. Moreover, this method is inexpensive and is applicable to multifragment cloning.
Nagano
Yukio
Y
Analytical Research Center for Experimental Sciences, Saga University, Saga, Japan. nagano@cc.saga-u.ac.jp
Takao
Syoko
S
Kudo
Takahiro
T
Iizasa
Ei'ichi
E
Anai
Toyoaki
T
D003001:Cloning, Molecular / Q000379:methods; D004269:DNA, Bacterial / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008957:Models, Genetic; D012275:Oryza / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D014170:Transformation, Genetic / Q000235:genetics*; D015003:Yeasts / Q000235:genetics*
D004269:DNA, Bacterial; C036483:T-DNA; D049452:Green Fluorescent Proteins