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Title: Cloning, expression and antimicrobial activity of crustinPm1, a major isoform of crustin, from the black tiger shrimp Penaeus monodon
Authors: Premruethai Supungul
Sureerat Tang
Cherdsak Maneeruttanarungroj
Vichien Rimphanitchayakit
Ikuo Hirono
Takashi Aoki
Anchalee Tassanakajon
Mahidol University
Thailand National Center for Genetic Engineering and Biotechnology
National University Corporation Tokyo University of Marine Science and Technology
Keywords: Biochemistry, Genetics and Molecular Biology;Immunology and Microbiology
Issue Date: 1-Jan-2008
Citation: Developmental and Comparative Immunology. Vol.32, No.1 (2008), 61-70
Abstract: Crustin antibacterial homologues, containing a whey acidic protein (WAP) domain, have been identified from the haemocyte library of the black tiger shrimp, Penaeus monodon. Sequence analysis of these cDNAs indicates the presence of several isoforms of crustin in P. monodon. CrustinPm1, the most abundant isoform, contains an open reading frame of 435 bp encoding a precursor of 145 amino acids that comprises 17 amino acid signal peptides and 128 amino acid mature peptides. The peptides contain a Gly-Pro rich region at the amino-terminus and a single whey acidic protein (WAP) domain at the carboxyl-terminus. In order to characterize the properties and biological activities of this peptide, crustinPm1 was overexpressed in Escherichia coli. The recombinant crustinPm1 has a molecular mass of 14.7 kDa with a predicted pI of 8.3. Antimicrobial assays demonstrated that recombinant crustinPm1 exhibited antimicrobial activity against only Gram-positive bacteria with strong inhibition against Staphylococcus aureus and Streptococcus iniae. In addition, the study of inhibition mechanism revealed that the antimicrobial activity of crustinPm1 was a result of bactericidal effect. In situ hybridization with crustinPm1 antisense probes showed strong hybridization signals in a certain haemocyte population of unchallenged shrimp, indicating that crustinPm1 transcript is differentially expressed in different subsets of haemocyte cells. © 2007 Elsevier Ltd. All rights reserved.
ISSN: 0145305X
Appears in Collections:Scopus 2006-2010

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