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Bombyx gene promoter analysis in transplanted silk gland transformed by particle delivery system
Author(s) -
Horard B.,
Mangé A.,
Pelissier B.,
Couble P.
Publication year - 1994
Publication title -
insect molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.955
H-Index - 93
eISSN - 1365-2583
pISSN - 0962-1075
DOI - 10.1111/j.1365-2583.1994.tb00175.x
Subject(s) - bombyx mori , biology , transgene , hemolymph , gene delivery , gene , bombyx , transfection , microbiology and biotechnology , promoter , dna , gene expression , genetics , botany
In most, if not all non‐Drosophila insect systems, the study of gene regulation relies on in vitro analyses. With the aim of providing an in vivo assay, we developed an original protocol using the Bombyx mori silk gland, by which the functioning of cloned gene promoters can be efficiently measured. This consists of introducing DNA‐coated tungsten particles into isolated silk glands by ballistic methods, according to a modified protocol in which the DNA is maintained in a hydrated state. The treated organ is then grafted into the haemolymph cavity of a host larva before analysis of transgene expression. In these reconstituted physiological conditions more than half of the cells express the transfected gene construct, and expression is both promoter‐dependent and cell type‐specific.