Application of Gibson Assembly method for plant expression vector construction

【Objective】 To construct a convenient and efficient plant expression vectors.【Method】 Gibson et al developed an isothermal in vitro recombination system or Gibson Assembly for ligation of multiple DNA fragments. The primers were designed to amplify target DNA sequences that contained 20-25 bp overlapping ends. The target DNA segments， linearized vector and the enzyme mixture were mixed in one tube to perform the assembly reaction.【Result and conclusion】 Using this method, several vectors for expressing rice genes were assembled. To increase the efficiency for cloning multiple fragments, the assembled primary products by PCR were amplified and then ligated into the vector. The Gibson Assembly method is faster and simpler without the limitation of internal restriction endonuclease sites of target genes, and it can be applied widely to construction of various vectors.