Hemolysin and its protein fragments in sero-detection of anaplasma phagocytophilum
Disclosed is the cloning, expression and purification of a hemolysin protein and its protein fragments in Anaplasma phagocytophilum . The recombinant hemolysin and its protein fragments are useful in the ELISA detection of anaplasma pathogen. The use of same as a kit for ELISA is also disclosed.
1. A composition comprising an isolated polypeptide and a support, wherein said isolated polypeptide is a recombinant polypeptide, consisting of the amino acid sequence selected from the group consisting of SEQ ID NO: 19, SEQ ID NO: 21, and SEQ ID NO: 23.
2. The composition of claim 1 , wherein said isolated recombinant polypeptide consists of the amino acid sequence set forth in SEQ ID NO: 19.
3. The composition of claim 1 , wherein said isolated recombinant polypeptide consists of the amino acid sequence set forth in SEQ ID NO: 21.
4. The composition of claim 1 , wherein said isolated recombinant polypeptide consists of the amino acid sequence set forth in SEQ ID NO: 23.
5. The composition of claim 1 , wherein said support is selected from the group consisting of polyethylene, polypropylene and glass.
6. The composition of claim 1 , wherein said support is a microtiter well.
7. The composition of claim 2 , wherein said isolated polypeptide is recombinantly expressed using an isolated polynucleotide, said isolated polynucleotide has a nucleotide sequence set forth in SEQ ID NO: 18.
8. The composition of claim 3 , wherein said isolated polypeptide is recombinantly expressed using an isolated polynucleotide, said isolated polynucleotide has a nucleotide sequence set forth in SEQ ID NO: 20.
9. The composition of claim 4 , wherein said isolated polypeptide is recombinantly expressed using an isolated polynucleotide, said isolated polynucleotide has a nucleotide sequence set forth in SEQ ID NO: 22.
10. A method of preparing the composition of claim 2 , comprising the steps of:
(i) introducing an isolated polynucleotide into a host cell, said isolated polynucleotide has a nucleotide sequence set forth in SEQ ID NO: 18;
(ii) growing said host cell in a culture under suitable conditions to permit production of said isolated polypeptide;
(iii) isolating said isolated polypeptide, and
(iv) providing a support, wherein, said composition is comprised of said isolated polypeptide and said support.
11. A method of preparing the composition of claim 3 , comprising the steps of:
(i) introducing an isolated polynucleotide into a host cell, said isolated polynucleotide has a nucleotide sequence set forth SEQ ID NO: 20;
(ii) growing said host cell in a culture under suitable conditions to permit production of said isolated polypeptide;
(iii) isolating said isolated polypeptide, and
(iv) providing a support, wherein, said composition is comprised of said isolated polypeptide and said support.
12. A method of preparing the composition of claim 4 , comprising the steps of:
(i) introducing an isolated polynucleotide into a host cell, said isolated polynucleotide has a nucleotide sequence set forth in SEQ ID NO: 22;
(ii) growing said host cell in a culture under suitable conditions to permit production of said isolated polypeptide; and
(iii) isolating said isolated polypeptide, and
(iv) providing a support, wherein, said composition is comprised of said isolated polypeptide and said support.
13. The composition of claim 1 , further comprising an isolated polypeptide and a support, wherein said isolated polypeptide is a recombinant polypeptide consisting of the amino acid sequence set forth in SEQ ID NO: 17.
14. The composition of claim 13 , wherein said support is selected from the group consisting of polyethylene, polypropylene and glass.
15. The composition of claim 13 , wherein said support is a microtiter well.
16. The composition of claim 13 , wherein said isolated polypeptide set forth in SEQ ID NO: 17 is produced by a method comprising the steps of:
(i) introducing an isolated polynucleotide into a host cell, said isolated polynucleotide has a nucleotide sequence set forth in SEQ ID NO: 16;
(ii) growing said host cell in a culture under suitable conditions to permit production of said isolated polypeptide; and
(iii) isolating said isolated polypeptide.