In vitro transcription systems from cultured cells and fat-body tissue of the silkworm,Bombyx mori

Abstract

This article describes the development of cell-free transcription systems from the cultured cells and fat body tissues of a Lepidpteran insect, the silkworm, Bombyx mori. Detailed protocols are provided for the culture of a B. mori cell line, rearing larvae, preparation of whole cell as well as nuclear extract and conditions for in vitro transcription of cloned plasma protein gene templates.

Functional characterization of the vitellogenin promoter in the silkworm, Bombyx mori

“…The in vitro fat body culture was carried out based on previous reports for B. mori and Ae. aegypti (Mine et al ., ; Roy et al ., ). Fat bodies were isolated from the female larvae at the L5D3 stage, when haemolymph 20E levels were low and the fat body was sensitive to 20E.…”

A serine protease in the midgut of the silkworm, Bombyx mori: Protein sequencing, identification of cDNA, demonstration of its synthesis as zymogen form and activation during midgut remodeling

“…Eggs of a hybrid strain (Kinshu  Showa) of the silkworm (B. mori) were obtained from Ueda Sanshu, and the hatched larvae were reared on an artificial diet (Nihon Nosan Kogyo) at 25 C as described elsewhere (Mine et al, 1997).…”

The homeodomain protein PBX participates in JH-related suppressive regulation on the expression of major plasma protein genes in the silkworm, Bombyx mori

“…Nuclear extracts from larval fat body cells were prepared as described by Mine et al (1997). The 5 0 end-labeled probe DNA was prepared as follows.…”