Cloning and prokaryotic expression of Populus tomentosa Carr. 4CL-like gene

In order to clone and analyze the prokaryotic expression of 4CL-like gene from Populus tomentosa Carr., total RNA were isolated, and 4CL-like gene were cloned by PCR. Phylogenetic analysis of the 4CL-like gene compared with other nine 4CL genes form different species with MEGA. Then it was over -expressed in prokaryotic systems. The expression construct pET30 -4CL -like was preformed. Based on the results, the bioinformational analysis showed that the CDS region of the nucleotide sequence was 1 758 bp, which could code 585 predicted aminoacids. Phylogenetic analysis showed that this gene was most similar to 4CL-like5 in Vitis vinifera. The SDS-PAGE analysis showed that the 4CL-like gene was transformed into E. coli BL21 and expressed successfully, fusion protein with molecular weight of 60 ku was successfully expressed in transformant.