title: Molecular cloning and expression analysis of a hemolin-like molecule from Antheraea pernyi.
abstract: 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 1418bp 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.
authors: College of Life Sciences, Anhui Agricultural University, 130 Changjiang West Road, Hefei 230036, China.
authors: Yu
authors: Y
authors: Sun
authors: College of Life Sciences, Anhui Agricultural University, 130 Changjiang West Road, Hefei 230036, China.
authors: Lei
authors: L
authors: Wang
authors: College of Life Sciences, Anhui Agricultural University, 130 Changjiang West Road, Hefei 230036, China.
authors: Cen
authors: C
authors: Qian
authors: College of Life Sciences, Anhui Agricultural University, 130 Changjiang West Road, Hefei 230036, China.
authors: Lishang
authors: L
authors: Dai
authors: College of Life Sciences, Anhui Agricultural University, 130 Changjiang West Road, Hefei 230036, China.
authors: Jun
authors: J
authors: Li
authors: College of Life Sciences, Anhui Agricultural University, 130 Changjiang West Road, Hefei 230036, China.
authors: Congfen
authors: C
authors: Zhang
authors: College of Life Sciences, Anhui Agricultural University, 130 Changjiang West Road, Hefei 230036, China. Electronic address: zhubaojian@ahau.edu.cn.
authors: Bao-Jian
authors: BJ
authors: Zhu
authors: College of Life Sciences, Anhui Agricultural University, 130 Changjiang West Road, Hefei 230036, China. Electronic address: cyschx@163.com.
authors: Chao-Liang
authors: CL
authors: Liu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D017344:Genes, Insect; D006458:Hemolymph / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D007113:Immunity, Innate; D007136:Immunoglobulins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics / Q000276:immunology*; D010802:Phylogeny; D060888:Real-Time Polymerase Chain Reaction; D011994:Recombinant Proteins; D016415:Sequence Alignment
chemical_list: D018076:DNA, Complementary; D007136:Immunoglobulins; D019476:Insect Proteins; D011994:Recombinant Proteins; C066846:hemolin
keywords: Antheraea pernyi; Expression pattern; Hemolin-like; Immune response