Abstract
Tools that allow for rapid, accurate and inexpensive assembly of multi-component combinatorial libraries of DNA for transformation into plants will accelerate the progress of synthetic biology research. Recent innovations in molecular cloning methods has vastly expanded the repertoire with which plant biologists can engineer a transgene. Here we describe a new set of binary vectors for use in Agrobacterium-mediated plant transformation that utilizes the Golden-Gate Cloning approach. Our optimized protocol facilitates the rapid and inexpensive generation of multi-component transgenes for later introduction into plants.
Highlights
Assembly and transformation of a multi-component DNA construct such as a promoter and a reporter gene fusion is a common task in everyday plant research
Frontiers in Plant Science | Plant Systems Biology or herbicides that may have a negative effect on normal growth and development, and T1 plants can be used for analysis
The Pro35S:PM-mCherry reporter is useful at later stages of transgenic line characterization for scoring the segregation ratio of the transgene
Summary
And transformation of a multi-component DNA construct such as a promoter and a reporter gene fusion is a common task in everyday plant research. Optimization of this process is likely to yield major productivity gains. Restriction endonuclease-mediated cleavage in combination with T4 DNA ligase-mediated joining has been used to create the desired DNA construct. This method is time consuming, sequence dependent and is not well suited for high-throughput assembly of a large number of constructs. Gateway cloning has significant drawbacks, which include the high cost of cloning kits, the need to use specific bacterial strains to propagate plasmids carrying the ccdB gene and the presence of recombination scars in the final product that can have effects on gene expression or protein function
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
