Direct analysis of RNA transcripts in electroporated carrot protoplasts
โ Scribed by Elizabeth E. Murray; Wallace G. Buchholz; Benjamin Bowen
- Book ID
- 104674848
- Publisher
- Springer
- Year
- 1990
- Tongue
- English
- Weight
- 970 KB
- Volume
- 9
- Category
- Article
- ISSN
- 0721-7714
No coin nor oath required. For personal study only.
โฆ Synopsis
We describe a method for direct analysis of RNA transcribed from DNA introduced into carrot cells by electroporation. Octopine synthase RNA transcribed from the plasmid p35SOcs was detected in total and poly A(+) RNA on Northern blots and in RNA protection assays. The highest level of octopine synthase transcript was detected at approximately 8 hrs post-electroporation, although RNA could still be detected after 48 hrs. This method allows detection of foreign gene expression in a plant system and bypasses the need for reporter genes.
๐ SIMILAR VOLUMES
We have investigated the basis for increased transient reporter gene expression following electroporation of protoplasts from uniform carrot cell suspension cultures at increasing DNA concentrations. Use of a combination of histochemical and fluorometric GUS gene assays allowed differentiation betwe
We have developed a simplified procedure for the detection of cellular RNA transcripts. Entire protoplasts, isolated nuclei or nuclei sorted according to cell cycle phases are directly dotted onto filters and hybridized. Transcripts are quantified thereupon without any RNA extraction methodology. Pr