[Recombinant prokaryotic plasmid construction and high expression of FUS1 gene].
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.
Zhang
Bao
B
Central Laboratory1, Clinical Laboratory, Tumor Hospital2, Shantou University Medical College, Shantou 515041, China. zhengff2004@gmail.com
Huo
Xia
X
Peng
Lin
L
Qi
Zong-li
ZL
Xu
Xi-jin
XJ
Li
Yan
Y
Qiu
Bo
B
Zheng
Liang-kai
LK
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
D011994:Recombinant Proteins; C499993:TUSC2 protein, human; D025521:Tumor Suppressor Proteins