Isolation of a dual plant promoter fragment from the Ti plasmid of Agrobacterium tumefaciens

Abstract

The two most abundant transcripts derived from TR‐DNA within plant cells transformed by an octopine strain of Agrobacterium tumefaciens arise from divergent transcription, both originating within an ˜500 bp section of the T‐DNA. Using a combination of subcloning and exonuclease digestion, a 479‐bp DNA fragment, directly flanked by the initiation codons for the two adjacent open reading frames, was isolated. The resulting DNA fragment was fused, in both orientations, to the neomycin phosphotransferase (NPT II) gene of the transposon Tn5 prior to introduction into Nicotiana tabacum cells via the Ti plasmid. The intergenic fragment was found to initiate expression of the NPT II gene in either orientation as assayed by kanamycin resistance of the transformed plant tissue as well as by enzymatic assay of the NPT II gene product. The plasmids described here are potential selection‐expression vectors for plant systems.

A Single-Electron Reducing Quinone Oxidoreductase Is Necessary to Induce Haustorium Development in the Root Parasitic Plant Triphysaria

“…The mas2' promoter (Velten et al, 1984) was PCR amplified from pFGC5941 (Gendler et al, 2008) and inserted into the NcoI site at the start of the YFP reporter gene in pBIN-YFP (Subramanian et al, 2006). The mas-YFP fusion was then cloned into the SacI site of pHellsgate8 to allow visual screening of transgenic roots (Helliwell et al, 2002).…”

Altered Growth and Wood Characteristics in Transgenic Hybrid Aspen Expressing Agrobacterium tumefaciens T-DNA Indoleacetic Acid-Biosynthetic Genes

“…The bidirectional 1'2' promoter was originally isolated from the T,-DNA of the octopine-type Ti plasmid pTiAch5 (Velten et al, 1984), which allows simultaneous expression of two genes from the divergent 1' and 2' promoters (Velten and Schell, 1985). A promoter-truncated iaaH gene (Sitbon et al, 1992b) was linked with SalI-linkers and inserted in sense orientation into the unique Sal1 site of the pPCV720 vector on the 3' side of the 2' promoter and upstream of a T-DNA octopine synthase poly(A) signal.…”

Aberrant Transpositions of Maize Double Ds-Like Elements Usually Involve Ds Ends on Sister Chromatids.

“…pSLJ4303 was based on pJJ4048 , a binary vector carrying a hygromycin phosphotransferase gene (Gritz and Davies, 1983) under the control of the Agrobacterium tumefaciens 1' promoter (Velten et al, 1984) and a promoterless streptomycin phosphotransferase (SPT) gene (Maliga et al, 1988;Jones et al, 1989). A bglucuronidase (GUS) coding sequence (Jefferson et al, 1987) and Agrobacterium nopaline synthase 3'end (Depicker et al, 1982) were inserted downstream of an Agrobacterium 2' promoter (Velten et al, 1984) as a 2-kb Clal fragment. A phosphinothricin acetyltransferase coding sequence and 3'end (DeBlock et al, 1987) were inserted downstream of the cauliflower mosaic virus (CaMV) 35s promoter as a 2-kb Hindlll-Xhol fragment.…”