A transgenic silkworm expressing the immune-inducible cecropin B-GFP reporter gene.
To analyze cecropin B promoter (P-CecB) activity in vivo, we constructed transgenic silkworms that expressed EGFP under the control of P-CecB using the piggyBac transposable element. Genomic Southern blot analysis of the G1 and G2 generations indicated the stable insertion of EGFP in the genome. Injection of Escherichia coli cells into the larvae strongly induced EGFP expression in the fat bodies and all five hemocyte cell types. Northern blot analysis indicated that the expression kinetics of EGFP in the fat bodies following bacterial injection were correlated with that of endogenous CecB. Flow cytometric analysis of the hemocytes revealed that EGFP expression was increased by bacteria, but not by yeast. Our results indicate that the features of EGFP expression in the transgenic silkworm are equivalent to those of endogenous CecB and that P-CecB activation can be monitored by EGFP expression using transgenic silkworms.
Imamura
Morikazu
M
Insect Biotechnology and Sericology Department, National Institute of Agrobiological Sciences, Owashi 1-2, Tsukuba, Ibaraki, 305-8634, Japan.
Nakahara
Yuichi
Y
Kanda
Toshio
T
Tamura
Toshiki
T
Taniai
Kiyoko
K
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D015152:Blotting, Northern; D012831:Bombyx / Q000235:genetics*; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics; D005434:Flow Cytometry; D017930:Genes, Reporter; D005818:Genetic Engineering / Q000379:methods; D049452:Green Fluorescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D007113:Immunity, Innate; D019476:Insect Proteins / Q000235:genetics*; D007814:Larva / Q000033:anatomy & histology / Q000235:genetics / Q000276:immunology
D004251:DNA Transposable Elements; D019476:Insect Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C031162:cecropin B protein, Insecta