期刊
INTERNATIONAL IMMUNOPHARMACOLOGY
卷 26, 期 1, 页码 65-71出版社
ELSEVIER
DOI: 10.1016/j.intimp.2015.03.010
关键词
Antheraea pernyi; Hemolin-like; Expression pattern; Immune response
资金
- earmarked fund for modern Argoindustry Technology Research System [CARS-22 SYZ10]
- Anhui High Schools Natural Science Foundation [KJ2013B320]
- National 863 Plans Projects of China [2011AA100306]
- Sericulture Biotechnology Innovation Team [2013xkdt-05]
- Ph.D. Programs in Biochemistry and Molecular Biology [xk2013042]
- Natural Science Foundation of the Anhui Higher Education Institutions of China [KJ2014A071]
Hemolin belongs to the immunoglobulin superfamily and plays an important role in innate immune response of insects. In this study, a hemolin-like cDNA of 1418 bp was obtained from Antheraea pernyi (Ap-hemolin-like). Sequence analysis revealed Ap-hemolin-like was homologous to those hemolins from other insect species. Recombinant Ap-hemolin-like protein was expressed in Escherichia coli cells, and polyclonal antibodies were produced against the recombinant proteins. Real-time PCR and western blot analysis showed that the Ap-hemolin-like was expressed in hemolymph, Malpighian tubules, midgut, epidermis and fat body, with the highest expression level in hemolymph. To investigate its role in the immune response against microorganisms, fifth instar larvae were challenged by injecting nucleopolyhedrovirus (NPV), E. coli, or Beauveria bassiana. The results showed that the expression of Ap-hemolin-like in hemolymph and fat body was obviously-induced by microorganisms. In addition, the recombinant Ap-hemolin-like protein promoted the agglutination of E. coli in the presence of calcium, which was confirmed by agglutination assay. These results suggested that the Ap-hemolin-like protein was involved in innate immune response of A. pernyi against pathogens. (C) 2015 Elsevier B.V. All rights reserved.
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