Sequence variation in trypsin- and chymotrypsin-like cDNAs from the midgut of Ostrinia nubilalis: methods for allelic differentiation of candidate Bacillus thuringiensis resistance genes

Midgut expressed alkaline serine proteases of Lepidoptera function in conversion of Bacillus thuringiensis (Bt) protoxin to active toxin, and reduced level of transcript T23 is associated with Ostrinia nubilalis resistance to Dipel® Bt formulations. Three groups of trypsin- (OnT25, OnT23, and OnT3)...

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Bibliographic Details
Published in:Insect molecular biology 2006-02, Vol.15 (1), p.13-24
Main Authors: Coates, B.S, Hellmich, R.L, Lewis, L.C
Format: Article
Language:English
Subjects:
Alleles
Amino Acid Sequence
amino acid sequences
Animals
Bacillus thuringiensis
Bacillus thuringiensis - physiology
Bacillus thuringiensis resistance
biological control agents
chymotrypsin
Chymotrypsin - genetics
complementary DNA
disease resistance
DNA, Complementary
entomopathogenic bacteria
Female
Gastrointestinal Tract - enzymology
genetic variation
Genome, Insect
insect pests
Lepidoptera
Male
Molecular Sequence Data
Moths - enzymology
Moths - genetics
Moths - microbiology
Ostrinia nubilalis
plant pests
Polymorphism, Single Nucleotide
Sequence Alignment
sequence analysis
Sequence Analysis, DNA
Serine Endopeptidases - genetics
serine proteases
Transcription, Genetic
Trypsin - genetics
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Summary:Midgut expressed alkaline serine proteases of Lepidoptera function in conversion of Bacillus thuringiensis (Bt) protoxin to active toxin, and reduced level of transcript T23 is associated with Ostrinia nubilalis resistance to Dipel® Bt formulations. Three groups of trypsin- (OnT25, OnT23, and OnT3) and two chymotrypsin-like (OnC1 and OnC2) cDNAs were isolated from O. nubilalis midgut tissue. Intraspecific groupings are based on cDNA similarity and peptide phylogeny. Derived serine proteases showed a catalytic triad (His, Asp, and Ser; except transcript OnT23a), three substrate specificity-determining residues, and three paired disulphide bonds. RT-PCR indicated all transcripts are expressed in the midgut. Mendelian-inherited genomic markers for loci OnT23, OnT3 and OnC1 will be useful for association of alleles with bioassayed Bt toxin resistance phenotypes.
ISSN:0962-1075
1365-2583
DOI:10.1111/j.1365-2583.2006.00598.x