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Expression of Secreted Metallothionein of Sinopotamon henanense Using Alkaline Phosphatase Promoter (phoA) and Signal Sequence in Escherichia coli

LIU Jin-ping, HE Yong-ji, WANG Lan   

  1. College of Life Science, Shanxi University, Taiyuan 030006, China
  • Online:2014-09-15 Published:2014-09-12

Abstract:

In this research we aimed to obtain soluble metallothionein (MT) of Sinopotamon henanense. Therefore, a
recombinant phoA plasmid containing MT open reading frame fused with a 6×His tag was constructed. The plasmid
possessed the promoter and signal sequence of the alkaline phosphatase gene of Escherichia coli. Subsequently, the
recombinant phoA-MT vector was transformed into E. coli BL21 (DE3) strain for MT expression. The fusion protein
induced by a low concentration of phosphate was purified by Ni2+ affinity chromatography and analyzed by UV absorption
spectrometry, SDS-PAGE and Western blotting. The results demonstrated that fusion protein was expressed with high purity
as a monomer and a dimmer, with molecular weight of approximately 7.5 and 15 kD, respectively. 

Key words: Sinopotamon henanense, metallothionein, expression

CLC Number: