Composition for preventing and treating
Disclosed is a composition for preventing and treating a Mycoplasma hyorhinis infection in swine. The composition uses XylF, DnaK, P72, or a combination thereof as an active pharmaceutical ingredient. Further disclosed are an expression vector and a method for producing the active pharmaceutical ingredient of the composition using a prokaryotic expression system.
1. A composition for avoiding a disease caused by Mycoplasma hyorhinis infection, comprising
an active ingredient comprising XylF, DnaK, P72 or a combination thereof; and
an adjuvant;
wherein said XylF comprises SEQ ID NO: 01, said DnaK comprises SEQ ID NO: 02, and said P72 comprises SEQ ID NO: 03;
wherein said disease is at least one selected from peritonitis, pleurisy, pericarditis and joint swelling.
2. The composition of claim 1 , wherein said active ingredient comprises at least two selected from a group consisting of XylF, DnaK and P72.
3. The composition of claim 2 , wherein said active ingredient comprises a combination of XylF, DnaK and P72.
4. The composition of claim 1 , wherein said active ingredient has a concentration of 50 to 300 μg/mL, based on the total volume of said composition.
5. The composition of claim 1 , wherein said adjuvant comprises complete Freund's adjuvant, incomplete Freund's adjuvant, alumina gel, surfactant, anionic polymer, peptide, oily emulsion, or a combination thereof.
6. The composition of claim 1 , wherein said disease is peritonitis, pleurisy or a combination thereof provided that said active ingredient is XylF.
7. The composition of claim 1 , wherein said disease is peritonitis provided that said active ingredient is DnaK.
8. The composition of claim 1 , wherein said disease is pleurisy provided that said active ingredient is P72.
9. The composition of claim 1 , wherein said disease is peritonitis, pleurisy, pericarditis and joint swelling provided that said active ingredient is a combination of XylF, DnaK and P72.
10. An expression vector for production of the active ingredient of the composition of claim 1 in a prokaryotic expression system, comprising:
an expression element, comprising a promoter and a ribosome binding site;
a nucleotide sequence encoding said XylF, said DnaK, said P72, or a combination thereof; and
a sequence encoding a fusion partner;
wherein said nucleotide sequence comprises SEQ ID NO: 04, SEQ ID NO: 05 or SEQ ID NO: 06, or a combination thereof.
11. The expression vector of claim 10 , wherein said fusion partner is DsbC of E. coli , MsyB of E. coli , FklB of E. coli , or a combination thereof.
12. The expression vector of claim 11 , wherein
said fusion partner is DsbC of E. coli provided that said nucleotide sequence encodes XylF;
said fusion partner is MsyB of E. coli provided that said nucleotide sequence encodes DnaK; or
said fusion partner is FklB of E. coli provided that said nucleotide sequence encodes P72.
13. The expression vector of claim 10 , further comprising a sequence encoding histidine tag, glutathione S-transferase tag or a combination thereof.
14. The expression vector of claim 10 , comprising a nucleotide sequence as shown in SEQ ID NO: 07, SEQ ID NO: 08, or SEQ ID NO: 09.
15. The expression vector of claim 10 , wherein said prokaryotic expression system is E. coli expression system.
16. A method for production of a soluble protein, wherein said protein is XylF, DnaK, P72, or a combination thereof; wherein said method comprising
providing a prokaryotic expression system; and
expressing said nucleotide sequence of the expression vector of claim 10 in said prokaryotic expression system.
17. The method of claim 16 , further comprising a step of passing a product obtained in the above step (2) through a nickel ion affinity column or a glutathione affinity column to obtain said soluble protein.