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Cloning and characterization of protease inhibitor‐like cDNAs from the Hessian fly mayetiola destructor (SAY)
Author(s) -
Maddur A. A.,
Liu X.,
Zhu Y. C.,
Fellers J. P.,
Oppert B.,
Park Y.,
Bai J.,
Wilde G. E.,
Chen M.S.
Publication year - 2006
Publication title -
insect molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.955
H-Index - 93
eISSN - 1365-2583
pISSN - 0962-1075
DOI - 10.1111/j.1365-2583.2006.00660.x
Subject(s) - biology , recombinant dna , gene , cloning (programming) , open reading frame , phylogenetic tree , orfs , gene family , accession number (library science) , genetics , sequence analysis , sequence alignment , genbank , peptide sequence , microbiology and biotechnology , biochemistry , gene expression , computer science , programming language
Analysis of transcriptomes from the salivary glands and midgut of Hessian fly larvae Mayetiola destructor (say) identified a set of diverse cDNAs that encode proteins with a relatively high percentage (over 10%) of cysteinyl residues. Structural comparison of these putative proteins with known sequences in G en B ank revealed that the positions of the cysteinyl residues in the identified proteins were highly conserved within a family of proteinase inhibitors despite very little overall sequence similarity. Phylogenetic analysis sorted this set of cDNAs into five different groups. To determine if these cDNAs indeed encode proteinase inhibitors, recombinant proteins were generated with two cDNAs from two different groups. Biochemical analysis of the recombinant proteins against commercial and insect gut proteinases demonstrated that the recombinant proteins are strong proteinase inhibitors with different specificities. Northern blot and real‐time PCR analysis revealed that the different genes were expressed at different developmental stages and in different tissues. The overall results indicated that M. destructor contains a complex of genes that code for proteinase inhibitors which may regulate proteinase activities in different regulatory pathways. The G en B ank accession numbers for the cDNAs in this paper were DQ232690 to DQ232718 .