Cauliflower mosaic virus 35S promoter

Fang, R.X., Nagy, E, Sivasubramaniam, S., and Chua, N.H. (1989). Multiple cis regulatory elements for maximal expression of the cauliflower mosaic virus 35S promoter in transgenic plants. Plant Cell 1(1) 141-150. 

Benfey, P.N. Chua, N.-H. (1990). The cauliflower mosaic virus 35S promoter combinatorial regulation of transcription in plants. Science 250, 959-66. 

Hex motif are found in the octopine synthase (ocs) and nopaline synthase (nos) promoters of the Ti-plasmids (Bouchez et al., 1989), as well as in the cauliflower mosaic virus 35S promoter (as-1 motif) (Katagiri et al.,... 

To test the hypothesis that phosphate supply from the can limit the rate of photosynthesis (Sivak and Walker, 1986), antisense experiments were performed by Schultz et al. (1993). A cDNA for the potato triose phosphate translocator was identified and a fragment of this cDNA in reverse orientation was expressed in trangenic potato plants under the control of the constitutive cauliflower mosaic virus 35S promoter (Rismeier et al., 1993). This experiment confirmed that Pi supply can limit photosynthesis since a... 

A -desaturase genes were isolated from cyanobacteria and Borago officinalis. Introduction of these genes into tobacco under the control of the cauliflower mosaic virus 35S promoter resulted in y-linolenic acid (GLA) production in leaf lipids. The use of seed specific promoters also resulted in significant levels of GLA in seed lipids. 

The identification of CPFA in callus triacylglycerol indicates that enzymes capable of dihydrosterculate transfer to that fraction are available in tobacco. The failure of the cauliflower mosaic virus 35S promoter to drive significant CPFA accumulation in seeds may therefore be unrelated to lipid metabolism perse. The 35S promoter is relatively weak during the latter part of seed development [9], and the relatively protease-sensitive CPFA synthase [10] may turn over rapidly. It is hoped that manipulation of promoters, enzyme localization, and substrate concentration will result in both dihydrosterculate-rich oils and a better understanding of fatty acid flow between phospholipid and triacylglycerol. 

Odell JT, Nagy F, Chua NH (1985) Identification of DNA sequences required for activity of cauliflower mosaic virus 35 S promoter. Nature 313 810-812... 

Fig. 7. Diagrams of the schemes for modifying levels of A, alcohol dehydrogenase and B, pyruvate decarboxylase activity and testing for survival of anoxia. In A, constructs contain the 35S promoter of the cauliflower mosaic virus (35S) driving expression of the cotton Adh cDNA in either the sense (Adh) or antisense (hdA) orientation, linked to the 3 termination signal of the nopaline synthase gene (Nos). Alternatively, the expression of cotton Adh cDNA is under control of the pea Adh promoter sequence (pea Adh). In B, either the 35S promoter or the pea Adh promoter is used to drive expression of the maize pyruvate decarboxylase cDNA (Pdc), linked to a Nos 3 termination sequence. Constructs are introduced into cotton via Agrobacterium tumefaciens-mediated infection of cotton. Transformed cotton callus is then assayed for its ability to survive anoxia.

The first hurdle encountered during the development of alfalfa as a recombinant protein production system was the relative inefficiency of the available expression cassettes. A study in which a tomato proteinase inhibitor I transgene was expressed in tobacco and alfalfa under the control of the cauliflower mosaic virus (CaMV) 35S promoter showed that 3-4 times more protein accumulated in tobacco leaves compared to alfalfa leaves [5]. Despite the low efficiency of the CaMV 35S promoter in alfalfa, bio-pharmaceutical production using this system has been reported in the scientific literature. Such reports include expression of the foot and mouth disease virus antigen [6], an enzyme to improve phosphorus utilization [7] and the anti-human IgG C5-1 [8]. In this last work, the C5-1 antibody accumulated to 1% total soluble protein [8]. 

As indicated in Table 2.1, most of the promoters used in plant tissue culture have been based on the constitutive cauliflower mosaic virus (CaMV) 35S promoter. In contrast, inducible promoters have the advantage of allowing foreign proteins to be expressed at a time that is most conducive to protein accumulation and stability. Although a considerable number of inducible promoters has been developed and used in plant culture applications, e.g. [32-37], the only one to be applied thus far for the production of biopharmaceutical proteins is the rice a-amylase promoter. This promoter controls the production of an a-amylase isozyme that is one of the most abundant proteins secreted from cultured rice cells after sucrose starvation. The rice a-amylase promoter has been used for expression of hGM-CSF [10], aranti-trypsin [12, 29, 38, 39] and human lysozyme [30]. 

Within each species, individual promoters resulted in distinct, tissue-dependent accumulation patterns. The cauliflower mosaic virus (CaMV) 35S promoter, for example, led to high-level accumulation in callus and leaves whereas the maize ubiqui-tin-1 promoter was the best choice for producing recombinant proteins in cereal seeds even though it is not in itself seed-specific [23]. The lack of such comparative studies for proteins other than rAbs makes it difficult to generalize an optimal expression strategy for all proteins. Tables 7.1 and 7.2 list recombinant proteins expressed in plants and provide details of the production system, promoters and other regulatory elements used in each case. 

The most common promoter element used in the design of transgenic plants is the 35S promoter of cauliflower mosaic virus (CaMV). This viral promoter is named for the sedimentation coefficient of its coordinating... 

Myb and bHLH protein-encoding genes have been ectopically expressed in the plant species from which they originated, as well as in heterologous plant species. All studies described here used the cauliflower mosaic virus (CaMV) 35S RNA promoter, which is highly active in most tissues of most plant species, to express the transcription factors. 

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