Functional analysis of four Gloverin-like genes in the silkworm, Bombyx mori.
To identify genes involved in the innate immunity of the silkworm Bombyx mori, we constructed a cDNA library from the fat body of Escherichia coli-challenged B. mori larvae. Based on the expressed sequence tag (EST) data and whole genome shotgun sequence analysis, we found four Gloverin-like genes, BmGlov1-4, in the Bombyx genome. Northern blot and RT-PCR analysis showed that BmGlov1-4 were induced in the larval fat body after an immune challenge by the injection of E. coli; however, less induction was observed after the injection of a yeast Candida albicans. In silico sequence analysis revealed the presence of a motif homologous to NF-kappaB binding site in the upstream region of each BmGlov gene. Moreover, we expressed recombinant BmGlov1-4 proteins using the baculovirus expression system, and found that all the recombinant BmGlov1-4 significantly inhibited the growth of E. coli.
Kawaoka
Shinpei
S
Department of Agricultural and Environmental Biology, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo, Japan.
Katsuma
Susumu
S
Daimon
Takaaki
T
Isono
Ryoko
R
Omuro
Naoko
N
Mita
Kazuei
K
Shimada
Toru
T
D000199:Actins / Q000032:analysis / Q000096:biosynthesis; D000595:Amino Acid Sequence; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis / Q000494:pharmacology; D001483:Base Sequence; D015152:Blotting, Northern; D012831:Bombyx / Q000235:genetics* / Q000276:immunology / Q000382:microbiology; D002176:Candida albicans / Q000276:immunology; D018076:DNA, Complementary / Q000737:chemistry; D004926:Escherichia coli / Q000187:drug effects / Q000276:immunology; D020224:Expressed Sequence Tags / Q000737:chemistry; D005216:Fat Body / Q000276:immunology / Q000378:metabolism; D005786:Gene Expression Regulation / Q000276:immunology*; D049750:Genome, Insect / Q000235:genetics; D036341:Intercellular Signaling Peptides and Proteins; D007814:Larva / Q000235:genetics / Q000276:immunology / Q000382:microbiology; D008969:Molecular Sequence Data; D010802:Phylogeny; D011401:Promoter Regions, Genetic / Q000235:genetics; D011506:Proteins / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D013997:Time Factors
D000199:Actins; D023181:Antimicrobial Cationic Peptides; D018076:DNA, Complementary; D036341:Intercellular Signaling Peptides and Proteins; D011506:Proteins; D011994:Recombinant Proteins; C107617:gloverin