オオムギ子葉鞘細胞における外来遺伝子の発現解析システム--マイクロインジェクションによる遺伝子の導入とin situ検出〔英文〕

概要

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The foreign genes, β-glucuronidase gene and TMV coat protein gene both ligated with the CaMV 35S promoter and nopalinsynthase terminator, were introduced into the inner epidermal cells of this tissue, using a barley coleoptile/microinjection system. The expression of the introduced genes was successfully examined by cytochemical identification of the transcription and translation products of the genes. Transcripts of the introduced genes were in situ hybridized to photobiotin conjugated antisense RNAs which were secondarily introduced by a pricking microinjection, and the translation products were detected by a complex formation with injected enzyme-conjugated antibodies. With a dual microinjection method, sense RNAs of these genes could be introduced and their translation products were detected at the same frequency (more than 80% of successfully injected cells) as those in DNA-injected cells. Thus, the present system, a combination of microinjection and in situ cytochemical techniques, allows introduction of foreign genes into barley coleoptile epidermal cells and precise detection of foreign gene expression in a higher plant cell system.
日本植物細胞分子生物学会の論文