Cloning and complete sequence characterization of two gypsy moth aminopeptidase-N cDNAs, including the receptor for Bacillus thuringiensis Cry1Ac toxin.
The complete cDNAs corresponding to two distinct gypsy moth (Lymantria dispar) larval gut aminopeptidases, APN1 and lambda APN2, were cloned and sequenced. The 3.4 kilobasepair cDNA of APN1 which encodes a 1017 amino acid prepro-protein corresponds to the previously-identified gypsy moth APN (APN-1) that specifically binds the Cry1Ac delta-endotoxin of Bacillus thuringiensis. Analysis of the primary structure of APN1 revealed a cluster of five potential N-linked glycosylation sites near the N-terminus and a C-terminal sequence characteristic of a putative glycosylphosphatidyl-inositol (GPI) anchor signal sequence. The cDNA of APN1 encodes the N-terminal peptide sequence and nine internal sequences obtained from the purified brush border membrane vesicle Cry1Ac receptor by protein sequencing. The lambda APN2 cDNA encodes a shorter protein with 51% similarity to APN1 that also appears to have a GPI anchor signal sequence. Expression of the APN1 cDNA in a baculovirus vector was confirmed by immunoblotting.
USDA Forest Service, Northeastern Research Station, Delaware, OH 43015, USA.
K J
KJ
Garner
S
S
Hiremath
K
K
Lehtoma
A P
AP
Valaitis
D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000235:genetics*; D000818:Animals; D001413:Bacillus thuringiensis; D001426:Bacterial Proteins; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D015342:DNA Probes; D018076:DNA, Complementary; D015870:Gene Expression; D005822:Genetic Vectors; D019476:Insect Proteins; D008969:Molecular Sequence Data; D009036:Moths / Q000201:enzymology* / Q000235:genetics; D017924:Nucleopolyhedroviruses; D011956:Receptors, Cell Surface / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D018411:Spodoptera / Q000166:cytology
D001426:Bacterial Proteins; C095416:Cry toxin receptors; D015342:DNA Probes; D018076:DNA, Complementary; D019476:Insect Proteins; D011956:Receptors, Cell Surface; D000626:Aminopeptidases