Tissue-specific expression from CaMV 35S enhancer subdomains in early stages of plant development.

Abstract

The cauliflower mosaic virus (CaMV) 35S enhancer is able to confer strong constitutive expression in plants. We have previously defined two domains within this enhancer that can confer different tissue‐specific expression patterns throughout development. We show here that the upstream domain (B) has a modular organization. It contains at least five subdomains that are able to confer distinct expression patterns when fused to the downstream domain (A). When fused to a minimal promoter only three of the five subdomains give any expression in the early stages of plant development. Comparison of the expression patterns conferred by the subdomains alone, in combination with the downstream domain or in combination with other subdomains provides evidence for synergistic interactions among cis‐elements within the 35S enhancer.

Characterisation of LTR sequences involved in the protoplast specific expression of the tobacco Tnt1 retrotransposon

“…Cell wall enzymatic digestions Constructs The 2120 plasmid (pBMCV 102120) (Fig 1) containing a translational LTR-GUS fusion (containing 110 bp of tobacco nitrate reductase genomic sequences, the entire Tntl 5' LTR, and the first 160 bp of the internal Tntl sequences) in a pBluescript KS plasmid has been previously described (9). The 2146 plasmid (pBMCV 102146), in which the LTR The EcoRI-BgllI fragment of the EMBLGUS46 plasmid (14), containing the minimal (-46,+8) CaMV 35S promoter fused to the reporter gene GUS and the 3C terminator from the pea rbc SE9 gene, was subcloned in the EcoRI-BamHI sites of a pUC18 plasmid, to give the construct pG46. The 300 bp HindIH fragment of the 2120 plasmid was cloned into the XbaI site of pG46, after filling-in with Klenow, to give the 4691 plasmid (pBMCV 104691) (Fig 2), in which the LTR sequence is in the same orientation as the 35S promoter, and the 4692 plasmid (pBMCV 104692) in which this sequence is in reverse orientation.…”

Different Temporal and Spatial Gene Expression Patterns Occur during Anther Development.

“…The -279 to +1 TA29 transcriptional control region ( Figure 13) was inserted directly into the pGV1500 GUS vector. By contrast, DNA fragments representing portions of this region were inserted into a similar GUS vector that contained the CaMV 35s gene minimal promoter (nucleotides -52 to +8; Benfey et al, 1990aBenfey et al, , 1990b. The chimeric TA29/DTA gene was constructed exactly as described by Mariani et al (1990) using a 5' TA29 gene fragment containing nucleotides -1 477 to +51 (Figure 1 OA) and the DTA coding sequence (Greenfield et al, 1983;Maxwell et al, 1986;Palmiter et al, 1987).…”

Light‐induced transcriptional repression of the pea AS1 gene: identification of cis‐elements and transfactors

“…The control construct, PB41 has four copies of the B4 subdomain of CaMV (-301 to -208) plus the A subdomain(-90 to +8) fused to GUS and was generated as described in Benfey et al (1990).…”

“…As controls, a promoterless construct (NN1019) and a constitutive promoter (PB41) were analysed (see Figure ld). PB41 transgenic tobacco seedlings contain the construct 35S CaMV(4XB4 +A)-GUS, which has four copies of the 35S CaMV promoter subdomain B4 (-301 to -208) plus subdomain A (-90 to +8) fused to GUS (Benfey et aL, 1990). This PB41 construct was previously shown to have a GUS expression pattern in the vascular tissue of the hypocotyl and cotyledons of 10-day old seedlings (Benfey etaL, 1990).…”

“…NN1019 is a promoterless-GUS construct. The control consruct, PB41 has four copies of the subdomain B4 (-301 to -208 of the 35S CaMV promoter) plus A (-90 to +8) fused to GUS and was generated as described in Benfey et al (1990). The gain-of-function construct, AS1-PB41 was created by inserting a PCR generated AS1 promoter from nt -124 to -33 into a Hindlll site just upstream to the PB41 element.…”