BR  >> Vol. 5 No. 2 (March 2016)

    Cloning and Prokaryotic Expression of LAC-1 Gene from Populus trichocarpa Torr. & Gray

  • 全文下载: PDF(1278KB) HTML   XML   PP.39-46   DOI: 10.12677/BR.2016.52007  
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毛果杨LAC基因克隆序列分析原核表达Populus trichocarpa LAC Gene Clone Sequence Analysis Prokaryotic Expression



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 expression vector to provide a foundation for studying the function of the target protein. Method: According 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 transformed into E. coli by the individual expression construct pET-30a and expression the recombination 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 SDS-PAGE demonstrated that the expressed proteins were consistent with the size of expected protein in the prokaryotic expression system. Conclusion: The LAC-1 genes belonged to the family of laccase and successfully expressed in E. coli. This study will supply theoretical foundation in identifying and functional analysis of members from laccase family.

李丽红, 撖静宜, 王莹, 曹山, 张强, 蒋璐瑶, 要笑云, 李慧, 陆海. 毛果杨漆酶基因LAC-1的克隆与原核表达研究[J]. 植物学研究, 2016, 5(2): 39-46.


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