title: [Recombinant prokaryotic plasmid construction and high expression of FUS1 gene].
abstract: OBJECTIVE
  To construct the prokaryotic plasmid of FUS1 gene for efficient FUS1 expression in E.coli strain Rosetta(DE3)2plys.
  
  
  METHODS
  The full-length FUS1 gene was amplified by PCR from the total RNA of umbilical mesenchymal stem cells and cloned into pET-32a(+) vector followed by identification with PCR and sequencing. The recombinant plasmid pET-32a(+)-FUS1 was transformed into the E.coli strain Rosetta(DE3)2plys and the target protein expression was induced by IPTG.
  
  
  RESULTS
  The plasmid pET-32a(+)-FUS1 was obtained successfully as verified by PCR and sequence analysis. High expression of the fused FUS1 protein was achieved after induction by low-concentration IPTG (25 micromol/L) for 3 h, and the recombinant FUS1 protein accounted for 40% of the total bacterial protein of Rosetta(DE3)2plys.
  
  
  CONCLUSIONS
  The recombinant FUS1 plasmid has been successfully cloned, which allows highly efficient FUS1 expression in Rosetta (DE3)2 plys.
authors.affiliation: Central Laboratory1, Clinical Laboratory, Tumor Hospital2, Shantou University Medical College, Shantou 515041, China. zhengff2004@gmail.com
authors.forename: Bao
authors.initials: B
authors.lastname: Zhang
authors.forename: Xia
authors.initials: X
authors.lastname: Huo
authors.forename: Lin
authors.initials: L
authors.lastname: Peng
authors.forename: Zong-li
authors.initials: ZL
authors.lastname: Qi
authors.forename: Xi-jin
authors.initials: XJ
authors.lastname: Xu
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Bo
authors.initials: B
authors.lastname: Qiu
authors.forename: Liang-kai
authors.initials: LK
authors.lastname: Zheng
mesh_terms: D015153:Blotting, Western; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D010957:Plasmids / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014170:Transformation, Genetic; D025521:Tumor Suppressor Proteins / Q000096:biosynthesis* / Q000235:genetics; D014470:Umbilical Cord / Q000166:cytology
chemical_list: D011994:Recombinant Proteins; C499993:TUSC2 protein, human; D025521:Tumor Suppressor Proteins