毛果杨漆酶基因LAC-1的克隆与原核表达研究.pdfVIP

Botanical Research 植物学研究, 2016, 5(2), 39-46 Published Online March 2016 in Hans. /journal/br /10.12677/br.2016.52007 Cloning and Prokaryotic Expression of LAC-1 Gene from Populus trichocarpa Torr. Gray Lihong Li, Jingyi Han, Ying Wang, Shan Cao, Qiang Zhang, Luyao Jiang, Xiaoyun Yao, Hui Li, Hai Lu College of Biological Science and Technology, Beijing Forestry University, Beijing th th th Received: Feb. 20 , 2016; accepted: Mar. 8 , 2016; published: Mar. 15 , 2016 Copyright © 2016 by authors and Hans Publishers Inc. This work is licensed under the Creative Commons Attribution International License (CC BY). /licenses/by/4.0/ Abstract Objective: Cloning, sequence analysis of the LAC-1 gene from Populus trichocarpa Torr. Gray, then induced expression and purify of the fusion protein in E. coli after construct prokaryotic expres- sion vector to provide a foundation for studying the function of the target protein. Method: Ac- cording to the principle of homologous cloning, laccase gene from Arabidopsis thaliana was used to blast the database JGI of Populus trichocarpa. The laccase gene was isolated by PCR and trans- formed into E. coli by the individual expression construct pET-30a and expression the recombina- tion protein, then purified by Ni-NTA affinity chromatography. Result: Populus trichocarpa laccase gene was isolated (renamed LAC-1, Genebank: XP_002310245). Seqence analysis revealed that LAC-1 had the key residues of laccase, phylogenetic analysis showed LAC-1 had high homologous with AtLAC4 . The results of SDSdemonstrated that the expressed proteins were consistent with the size of expected protein in the prokaryotic expression system. Conclusion: The