Simplification of Entry Vector by TA Approach

Gateway technology is a universal cloning approach that enables rapid cloning of DNA fragments into mul- tiple Gateway-compatible destination vectors using λ phage site-specific recombination , eliminating the requirement to work with restriction enzymes and ligase. But a problem using this system for making entry clone is the expensive- ness and long time to buy the enzyme. To solve this problem , we created the TA cloning entry vector that contained a T-tail in each 3 ' -end through modification of pDONR207. The TA cloning approach can construct entry clones simply , economically and rapidly. Using Gateway T vectors prepared by this improved method , prokaryotic expres- sion vector and eukaryotic expression vector for SOS2 gene were constructed. Through the methods of prokaryotic ex- pression and transient gene expression in Arabidopsis protoplasts , it proved that the SOS2 gene expressed well in both