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| The Construction and Functional Analysis of Staphylococcal Enterotoxin-like K |
| LIU Yan-li1,2, NIU Rong-cheng1, YANG Fen1, ZHU Wen-hui1, LIN Jun-tang1,2 |
1. College of Life Science and Technology, Xinxiang Medical University, Xinxiang 453003, China;
2. Henan Key Laboratory of Medical Tissue Regeneration, Xinxiang 453003, China |
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Abstract Objective:The aim is to construct the prokaryotic expression vector of Staphylococcal Enterotoxin-like K (SElK), and preliminarily determine its' biological activity. Methods:Firstly, the target gene SElK was cloned into the plasmid of pET-28a, after being confirmed by colony PCR, double digestion and sequence, the purified recombined protein SElK was achieved by Ni+-affinity chromatography, and examined by SDS-PAGE. Subsequently, the superantigen activity of SElK was examined by stimulating the proliferation of spleen lymphocytes in vitro, and the routine blood tests in vivo after SElK injection from tail vein. Finally, the intrinsic enterotoxigenesis of SElK was evaluated by examined the activity or content of key organs' important indicators in the serum after SElK injection from tail vein, including ALT, AST, LDH, CK, Urea and CR. Results:Prokaryotic expression vector of pET-28a-SElK was successfully constructed and high purity (>95%) recombined protein SElK was achieved by Ni+-affinity chromatography; the subsequent analysis of superantigen activity showed that SElK could significantly stimulate the proliferation of spleen lymphocytes in vitro, and the further routine blood tests also demonstrated that SElK could induce the expension of lymphocytes in vivo; the final enterotoxigenesis tests showed that SElK could significantly increase the activity of AST and the content of Urea. Conclusion:The provided fundamental data for the characteristics of SElK, and the results suggested that SElK could be used as a potent candidate agent for tumour treatment in clinics.
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Received: 13 October 2015
Published: 22 December 2015
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