A human metallothionein-3 fusion protein expression

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A human metallothionein-3 fusion protein expression vector is disclosed. The upstream of a cloning region of the expression vector comprises a human fatty acid binding protein (hFABP6) and the downstream of the cloning region comprises metallothionein MT3. A method of applying the expression vector is also disclosed and includes basic manners for expression strain construction, amplification, inducible expression and fusion protein purification. The hFABP6-MT3 fusion protein expression vector is characterized by (1) efficient expression of MT3 protein and (2) soluble expression.

The present invention belongs to the field of genetic engineering and fermentation engineering, and set up a human metallothionein -3 fusion protein expression vector, and in particular set up a kind of chaperone-like protein function of human free fatty acid binding protein and human metallothionein fusion expression.

1957 Margoshes and Vallee in the study of the biological effects of metal, isolated from animal organs of a new protein, it is rich in sulfur group can Xin together a large number of metal ions, this substance called metallothionein ( English as Me1: allothionein, called MT).

on metallothionein, a global From 1978 to 1999, worked in Switzerland ±, Japan, the United States held a total of four international symposia, the Fifth International Symposium in October 2005 was held in Beijing. 1987 China will be included in MT Program / 'eight five "," Nine Five "major research projects in 1994 included in the national; 2003 MT project by the National Science and Technology as" national scientific and technological achievements promote projects in 1995 included the United Nations to the world recommend MT biotechnology products. Thus from domestic to foreign countries for MT research, development, promotion and application of attention.

The object of the present invention is to provide a human metallothionein fusion protein expression vector.

The present invention may be realized by the following technical solutions:

Fusion kinds of human metallothionein fusion protein expression vector, the human metallothionein fusion protein expression vector is a human metallothionein -3 (MT3) coding sequence as a target protein coding sequence inserted chaperone-like proteins - [0011] the resulting expression vector cloning region; fusion protein expression vector upstream of said chaperone-like protein containing a sequence encoding the human free fatty acid binding protein-6, human free fatty acid binding protein coding sequence downstream of the zone and contains some flexible joint for insertion polyclonal target area.

The free fatty acid binding protein of the human and the natural human -6 free fatty acid binding protein homology equal or greater than 85%, of the free fatty acid binding protein -6 human coding sequence is preferably in the codon-optimized coding sequence.

The free fatty acid binding protein in human -6 coding sequence preferably as SEQ ID NO. 1 Fig.

The flexible linker region and polyclonal region sequences preferably as shown in SEQ ID NO.3.

The human metallothionein -3 amino acid sequence is preferably the amino acid sequence of natural human metallothionein homology equal or greater than 75%.

human metallothionein -3 (MT3) the amino acid sequence shown in SEQ ID NO.21, codon-optimized human metallothionein-3 (1 of 3) coding sequences are preferably as 56〇10 ^ .20 shown.

Upstream of the protein expression vector

The free fatty acids in human binding protein coding sequence comprises a coding-6 for further separation and purification tag fusion protein sequence, preferably a sequence encoding a histidine tag.

Cited beneficial effects:

1. The recombinant expression vectors of the present invention is constructed to be able to promote or induce fusion downstream human metallothionein fusion protein folding, protein having a chaperone-like characteristics.

2. The human protein FABP derived, in theory, there is no human immunogenicity, the pharmaceutical industry is suitable for the target protein inclusions, solve problems, and W can be retained with the fusion protein pharmaceuticals.