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Cloning and molecular analysis of the aspartic protease Sc-ASP110 gene transcript in Steinernema carpocapsae

Published online by Cambridge University Press:  04 June 2013

NATESAN BALASUBRAMANIAN  and
NELSON SIMÕES
NATESAN BALASUBRAMANIAN*
Affiliation:
CIRN and Department of Biology, University of Azores, 9501-855 Ponta Delgada, Portugal
NELSON SIMÕES
Affiliation:
CIRN and Department of Biology, University of Azores, 9501-855 Ponta Delgada, Portugal
*
*Corresponding author: CIRN and Department of Biology, University of Azores, 9501-855 Ponta Delgada, Portugal. E-mail: [email protected]; [email protected]
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Summary

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.

Keywords

Aspartic proteasedevelopmental stagegene transcriptparasitic nematodeSteinernema carpocapsae
Type
Research Article
Information
Parasitology , Volume 140 , Issue 9 , August 2013 , pp. 1158 - 1167
Copyright
Copyright © Cambridge University Press 2013 

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