Method and system for inducible recombinational cloning in bacterial cells

摘要

The present invention discloses methods and systems that enable high throughput screening of genes and DNA sequence bearing expressed sequence tags in mammalian cells. The invention can easily be applied to the delivery of other functions like that of RNAi, ribozyme, antisense, DNA vaccine, etc.. The invention comprises a donor plasmid bearing a donor cassette and a receiver plasmid bearing a recombinase cassette, which may be propagated and maintained in bacterial culture. Through the use of inducible recombinational cloning, the donor cassette comprising a transgene sequence and a receiver plasmid, are easily and efficiently recombined in vivo to create a target vector comprising desired motifs (e.g. splice sites, introns and polyadenylation sequences). The present invention also provides convenient means for the introduction of the transgene into the donor cassette and provides an ability to create a library of transgene sequences for study maintained either in the donor plasmid or in the target vector. Because of the ease with which genes and putative genes may be introduced into the donor plasmid, and that recombination occurs in vivo, the invention allows for the functional study of genomics in model systems.