Plant in Vitro Transcription Systems

Abstract

In vitro transcription systems provide a powerful tool for detailed analysis of transcription reactions including initiation, elongation, and termination. Despite problems inherent to plant cells, efforts have been made to develop plant in vitro transcription systems in the past decade. These efforts have finally culminated in the development of reliable in vitro systems from suspension-cell cultures of both monocot and dicot plants. These systems can be useful in elucidating the specific mechanisms involved in the process of plant transcription and thus can potentially open a new era of transcription studies in plants.

Efficient in vitro transcription of plant nuclear tRNASer genes in a nuclear extract from tobacco cultured cells

“…Plant in vitro systems have advantages for elucidating the specific and detailed mechanisms involved in the process and regulation of plant transcription (Sugiura, 1996;Zhu, 1996;Zhu et aL, 1995). Tobacco BY-2 nuclear extracts can be the versatile transcription system for Pol I, II and Ill-dependent plant genes in vitro (Sugiura, 1997), and tobacco is a good system for in vivo studies on gene expression using transient and transformed assays.…”

Isolation of putative plant transcriptional coactivators using a modified two‐hybrid system incorporating a GFP reporter gene

“…The knowledge of the processes of transcription in plants still lags considerably behind that of animals or yeast. One reason for this deficit is our rather limited success in establishing a convenient and reliable plant in vitro transcription system despite recent promising developments (Sugiura, 1997;Zhu, 1996). Nevertheless, due to the high level of conservation in the factors involved, the mechanism of transcriptional activation is believed to follow a common blueprint in all eukaryotic organisms (Stargell and Struhl, 1996).…”

Rice TATA Binding Protein Interacts Functionally with Transcription Factor IIB and the RF2a bZIP Transcriptional Activator in an Enhanced Plant in Vitro Transcription System

“…Analysis of RNA polymerase II-mediated transcription initiation in vitro is a powerful approach for the functional analysis of the transcription machinery and its regulation by specific cis element-trans factor interactions (Zhu, 1996;Sugiura, 1997). Efficient in vitro transcription by rice wholecell or tobacco nuclear extracts giving authentic initiation from the in vivo start site was reported recently (Fan and Sugiura, 1995; Zhu et al, 1995a).…”

“…This is followed by TFIIB binding to the TBP promoter complex to give a more stable ternary complex, which acts as the scaffold for RNA polymerase II and accessory factors (Nikolov et al, 1995;Orphanides et al, 1996;Tansey and Herr, 1997). However, although the plant RNA polymerase II transcription machinery appears to be very similar to that in other eukaryotes, little is known about the functional interactions underlying the selective transcription of plant genes and the mechanisms by which sequence-specific trans factors regulate the basal transcription machinery.Analysis of RNA polymerase II-mediated transcription initiation in vitro is a powerful approach for the functional analysis of the transcription machinery and its regulation by specific cis element-trans factor interactions (Zhu, 1996;Sugiura, 1997). Efficient in vitro transcription by rice wholecell or tobacco nuclear extracts giving authentic initiation from the in vivo start site was reported recently (Fan and Sugiura, 1995; Zhu et al, 1995a).…”