Purification, cDNA cloning and expression of an insect defensin from the great wax moth, Galleria mellonella.
An insect defensin, named Galleria defensin, was purified from the larval haemolymph of Galleria mellonella immunized against E. coli. The peptide was composed of forty-three amino acid residues containing six cysteines that might be engaged in intramolecular disulphide bridges. The primary structure of Galleria defensin shared about 90.7% identity to that of heliomicin, which was an insect defensin isolated from Heliothis virescens. The full-length cDNA encoding Galleria defensin was cloned from the fat body of the immunized G. mellonella larvae. Northern blot analysis revealed that Galleria defensin was expressed not only in the fat body but also in the midgut against invading bacteria into haemocoel. This is the first report presenting cDNA and expression of an insect defensin in the lepidopteran species.
Lee
Y S
YS
Department of Life Science, Hoseo University, South Korea.
Yun
E K
EK
Jang
W S
WS
Kim
I
I
Lee
J H
JH
Park
S Y
SY
Ryu
K S
KS
Seo
S J
SJ
Kim
C H
CH
Lee
I H
IH
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002851:Chromatography, High Pressure Liquid; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D023082:Defensins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005216:Fat Body / Q000378:metabolism; D015870:Gene Expression; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D018076:DNA, Complementary; D023082:Defensins