Clone and soluble fusion expression ofα-HL of Staphylococcus aureus
10.3969/j.issn.1000-484X.2016.04.018
- VernacularTitle:金黄色葡萄球菌α-溶血素的克隆及可溶性融合表达
- Author:
Tong WU
;
Wenfeng XU
;
Siji NIAN
;
Yingchun YE
;
Qing YUAN
- Publication Type:Journal Article
- Keywords:
Staphylococcus aureus;
α-HL;
Soluble expression
- From:
Chinese Journal of Immunology
2016;32(4):532-535,541
- CountryChina
- Language:Chinese
-
Abstract:
Objective:Expression and purification of the α-HL of Staphylococcus aureus as antigen for making full human anti-α-HL antibody later ,providing of new treatment for Staphylococcus aureus infection.Methods:The total RNA of Staphylococcus aureus was extracted and the cDNA of α-HL was amplified by RT-PCR.The DNA of α-HL and pCold-TF plasmid was digested and ligated by T4 ligase and then transformed into E.coli TOPO 10.The recombinant plasmid α-HL/pCold-TF which verified by sequencing was trans-formed into E.coli BL21 for expression.The expression products was identified by SDS-PAGE and Western blot.Results: The size of amplified cDNA of α-HL was about 900 bp and the expressed soluble fusion protein of α-HL was about 90 kD(including the molecular chaperone in the vector ) after inducing expression for 24 h at 15℃.The Western blot results showed that the expressed protein was the fusion protein of α-HL.The purified α-HL was injected into BABL/c mice for making antiserum.The results showed that the antiserum had good binding activity with Staphylococcus aureus and the titer was greater than 10 000 times.Conclusion: The α-HL of Staphylococcus aureus was successfully cloned and the soluble fusion protein of α-HL was successfully expressed.
