Preferences help
enabled [disable] Abstract
Number of results
2009 | 56 | 4 | 671-677
Article title

Molecular cloning, expression and characterization of Bmserpin-2 gene from Bombyx mori

Title variants
Languages of publication
Serpins are a broadly distributed family of protease inhibitors. In this study, the gene encoding Bombyx mori serpin-2 (Bmserpin-2) was cloned and expressed in E. coli. The Bmserpin-2 cDNA contains a 1125 bp open reading frame (ORF). The deduced protein has 374 amino-acid residues, contains a conserved SERPIN domain and shares extensive homology with other invertebrate serpins. RT-PCR analysis showed that Bmserpin-2 was expressed in all developmental stages of B. mori larvae and various larval tissues. Subcellular localization analysis indicated that Bmserpin-2 protein was located in the cytoplasm. Interestingly, real-time quantitative PCR revealed that the expression of Bmserpin-2 in the midgut of susceptible B. mori strain 306 significantly increased at 72 hours post inoculation (hpi) when infected with BmNPV. However, there was no significant increase of the Bmserpin-2 expression in resistant strain NB infected with BmNPV. Thus, our data indicates that Bmserpin-2 may be involved in B. mori antiviral response.
Physical description
  • Abraham EG, Pinto SB, Ghosh A, Vanlandingham DL, Budd A, Higgs S, Kafatos FC, Jacobs-Lorena M, Michel K (2005) An immune-responsive serpin, SRPN6, mediates mosquito defense against malaria parasites. Proc Natl Acad Sci USA 102: 16327-16332.
  • Chamankhah M, Braun L, Visal-Shah S, O'Grady M, Baldwin D, Shi X, Hemmingsen SM, Alting-Mees M, Hegedus DD (2003) Mamestra configurata serpin-1 homologues: cloning, localization and developmental regulation. Insect Biochem Mol Biol 33: 355-369.
  • Dementiev A, Dobó J, Gettins PGW (2006) Active site distortion is sufficient for proteinase inhibition by serpins: structure of the covalent complex of α1-proteinase inhibitor with porcine pancreatic elastase. J Biol Chem 281: 3452-3457.
  • Florea L, Hartzell G, Zhang Z, Rubin GM, Miller W (1998) A computer program for aligning a cDNA sequence with a genomic DNA sequence. Genome Res 8: 967-974.
  • Gan H, Wang Y, Jiang H, Mita K, Kanost MR (2001) A bacteria-induced, intracellular serpin in granular hemocytes of Manduca sexta. Insect Biochem Mol Biol 31: 887-898.
  • Hunt LT, Dayhoff MO (1980) A surprising new protein superfamily containing ovalbumin, antithrombin-III, and α1-proteinase inhibitor. Biochem Biophys Res Commun 95: 864-871.
  • Huntington JA, Read RJ, Carrell RW (2000) Structure of a serpin-protease complex shows inhibition by deformation. Nature 407: 923-926.
  • Irving JA, Pike RN, Lesk AM, Whisstock JC (2000) Phylogeny of the serpin superfamily: implications of patterns of amino acid conservation for structure and function. Genome Res 10: 1845-1864.
  • Jiang H, Kanost MR (1997) Characterization and functional analysis of 12 naturally occurring reactive site variants of serpin-1 from Manduca sexta. J Biol Chem 272: 1082-1087.
  • Kanost MR (1999) Serine proteinase inhibitors in arthropod immunity. Dev Comp Immunol 23: 291-301.
  • Khurad AM, Mahulikar A, Rathod MK, Rai MM, Kanginakudru S, Nagaraju J (2004) Vertical transmission of nucleopolyhedrovirus in the silkworm, Bombyx mori L. J Invertebr Pathol 87: 8-15.
  • Nakazawa H, Tsuneishi E, Ponnuvel KM, Furukawa S, Asaoka A, Tanaka H, Ishibashi J, Yamakawa M (2004) Antiviral activity of a serine protease from the digestive juice of Bombyx mori larvae against nucleopolyhedrovirus. Virology 321: 154-162.
  • Pike RN, Buckle AM, le Bonniec BF, Church FC (2005) Control of the coagulation system by serpins. Getting by with a little help from glycosaminoglycans. FEBS J 272: 4842-4851.
  • Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4: 406.
  • Salzet M, Vieau D, Stefano GB (1999) Serpins: an evolutionarily conserved survival strategy. Immunol Today 20: 541-544.
  • Silverman GA, Bird PI, Carrell RW (2001) The serpins are an expanding superfamily of structurally similar but functionally diverse proteins. Evolution, mechanism of inhibition, novel functions, and a revised nomenclature. J Biol Chem 276: 33293-33296.
  • Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24: 1596-1599.
  • Tong Y, Kanost MR (2005) Manduca sexta serpin-4 and serpin-5 inhibit the prophenol oxidase activation pathway: cDNA cloning, protein expression, and characterization. J Biol Chem 280: 14923-14931.
  • Van Gent D, Sharp P, Morgan K, Kalsheker N (2003) Serpins: structure, function and molecular evolution. Int J Biochem Cell Biol 35: 1536-1547.
  • Wong ML, Medrano JF (2005) Real-time PCR for mRNA quantitation. Biotechniques 39: 75-85.
  • Zhu Y, Wang Y, Gorman MJ, Jiang H, Kanost MR (2003) Manduca sexta serpin-3 regulates prophenoloxidase activation in response to infection by inhibiting prophenoloxidase-activating proteinases. J Biol Chem 278: 46556-46564.
Document Type
Publication order reference
YADDA identifier
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.