title: Cloning and molecular analysis of the aspartic protease Sc-ASP110 gene transcript in Steinernema carpocapsae.
abstract: Many protease genes have previously been shown to be involved in parasitism and in the development of Steinernema carpocapsae, including a gene predicted to encode an aspartic protease, Sc-ASP110, which was cloned and was analysed in this study. A cDNA encoding Sc-ASP110 was cloned based on an expressed sequence tag (EST) fragment from our EST library. The full-length cDNA of Sc-ASP110 consists of 1112 nucleotides with a catalytic aspartic domain (aa18-337). The putative 341 amino acid residues have a calculated molecular mass of 37·1 kDa and a theoretical pI of 4·7. BLASTp analysis of the Sc-ASP110 amino acid sequence showed 45-77% amino acid sequence identity to parasitic and non-parasitic nematode aspartic proteases. An expression analysis showed that the sc-asp110 gene was upregulated during the late parasitic stage, L4, and 24 h after induction of in vitro nematodes. A sequence comparison revealed that Sc-ASP110 was a member of an aspartic protease family; additionally, a phylogenetic analysis indicated that Sc-ASP110 was clustered with the closely related nematode Steinernema feltiae. In situ hybridization showed that sc-asp110 was expressed in the body walls of dorsal cells. The upregulated Sc-ASP110 expression revealed that this protease could play a role in the late parasitic process. In this study, we have cloned and analysed the gene transcript of Sc-ASP110 in S. carpocapsae.
authors.affiliation: CIRN and Department of Biology, University of Azores, 9501-855 Ponta Delgada, Portugal. natesanbala@uac.pt
authors.forename: Natesan
authors.initials: N
authors.lastname: Balasubramanian
authors.forename: Nelson
authors.initials: N
authors.lastname: Simões
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D057055:Aspartic Acid Proteases / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D019295:Computational Biology; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D018509:DNA, Helminth / Q000737:chemistry / Q000235:genetics; D005786:Gene Expression Regulation; D018507:Gene Expression Regulation, Developmental; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009036:Moths / Q000469:parasitology; D010802:Phylogeny; D018510:RNA, Helminth / Q000235:genetics / Q000302:isolation & purification; D012333:RNA, Messenger / Q000235:genetics; D017168:Rhabditida / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D015854:Up-Regulation
chemical_list: D018076:DNA, Complementary; D018509:DNA, Helminth; D018510:RNA, Helminth; D012333:RNA, Messenger; D057055:Aspartic Acid Proteases