Surface expression vector for fusion protein of Myo-2 peptide multimer and myostatin, and microorganism transformed by thereof
The present invention relates to a fusion protein in which a myostatin mature protein is fused to a multimer of myostatin-derived antigenic peptide Myo-2, a surface expression vector containing a polynucleotide encoding the fusion protein, a recombinant microorganism transformed with the vector, and a feedstuff additive or a pharmaceutical composition containing the microorganism as an effective ingredient. The feedstuff additive or pharmaceutical composition according to the present invention can be used for muscle development and regulation of muscle growth in livestock and poultry, as well as for preventing and treating muscle-wasting diseases and degenerative diseases such as muscular dystrophy, muscular atrophy and the like. In addition, the transformed strain shows the same effect even if the strain itself after culture thereof is directly used, and thus it is very economical.
1. A nucleic acid surface expression vector for expression of a myostatin fusion protein in a cell, comprising:
one or more than two genes encoding a poly-gamma-glutamate synthetase complex selected from the group consisting of pgsB, pgsC and pgsA, operatively linked to
a gene encoding a fusion protein comprising a myostatin mature protein fused with a Myo-2 peptide multimer consisting of 2 to 8 of an isolated Myo-2 peptide, wherein each Myo-2 peptide consist of the amino acid sequence of SEQ ID NO: 1.
2. The expression vector according to claim 1 , wherein said Myo-2 peptide is isolated from mammals or birds.
3. A recombinant bacterium which is obtained by transforming a bacterium selected from the group consisting of Gram-negative bacteria and Gram-positive bacteria with the expression vector according to claim 1 , wherein upon expression, the fusion protein is expressed on the cell surface.
4. The bacterium according to claim 3 , wherein the bacterium is selected from the group consisting of E. coli, Salmonella typhi, Salmonella typhimurium, Vibrio cholerae, Mycobacterium bovis, Shigella, Bacillus , lactic acid bacteria, Staphylococcus, Corynebacteria, Listeria monocytogenes and Streptococcus.
5. The bacterium according to claim 4 , wherein the bacterium is lactic acid bacteria.
6. An isolated Myo-2 peptide from myostatin consisting of the amino acid of sequence of SEQ ID NO: 1.
7. A Myo-2 peptide multimer consisting of 2 to 8 of the Myo-2 peptide of claim 6 .
8. A fusion protein comprising a myostatin mature protein fused with a Myo-2 peptide multimer consisting of 2 to 8 of an isolated Myo-2 peptide, wherein each Myo-2 peptide consist of the amino acid sequence of SEQ ID NO: 1.
9. A feedstuff additive for promoting muscle growth or improving body weight gain in animals, which comprises the fusion protein of claim 8 as an effective ingredient.
10. A composition for promoting muscle growth or improving body weight gain in animals, which comprises the fusion protein of claim 8 as an effective ingredient.