Cloning and analysis of a new 4CL-like gene in Populus tomentosa

The phenylpropanoid enzyme 4-coumarate:coenzyme A ligase (4CL), plays a key role in general phenylpropanoid metabolism. Our investigation involves a new 4CL-like gene cloned from Populus tomentosa. This new 4CL-like gene is 1,692 bp and encodes a protein of 552 aa. After analysis of its domain, we found a highly conserved region of a 4CL gene family in this 4CL-like gene. Based on our results, we believe that this gene belongs to the family of 4CL. A recombinant vector, referred to as pET30a-4CL-like, was constructed by connecting this 4CL-like gene fragment to pET30a. After inducing it with IPTG for 3 h, the 4CL-like protein was purified by a Ni-NTA method. This 4CL-like protein has a calculated molecular weight of 60 kDa by SDS-PAGE.