Expression of a soybean ?-conclycinin gene under the control of the Cauliflower Mosaic Virus 35S and 19S promoters in transformed petunia tissues

Lawton, Michael; Tierney, Mary A.; Nakamura, Ikuo; Anderson, E. J.; Komeda, Yoshi; Dubé, Philip; Hoffman, N; Fraley, Robert T.; Beachy, Roger N.

doi:10.1007/bf00014906

Cited by 60 publications

(21 citation statements)

References 32 publications

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“…This result is similar to that in an earlier report, where it was shown that the expression level in individual transformants varied even at identical gene copy number (24). The 35-kDa AmA1 protein was detected in both pSB8 and pSB8G immunoblots ( Fig.…”

Section: Immunodetection Of Ama1 Protein In Transgenic Tuberssupporting

confidence: 92%

Increased nutritive value of transgenic potato by expressing a nonallergenic seed albumin gene from Amaranthus hypochondriacus

Chakraborty

Chakraborty²,

Datta³

2000

Proceedings of the National Academy of Sciences

131

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Improvement of nutritive value of crop plants, in particular the amino acid composition, has been a major long-term goal of plant breeding programs. Toward this end, we reported earlier the cloning of the seed albumin gene AmA1 from Amaranthus hypochondriacus. The AmA1 protein is nonallergenic in nature and is rich in all essential amino acids, and the composition corresponds well with the World Health Organization standards for optimal human nutrition. In an attempt to improve the nutritional value of potato, the AmA1 coding sequence was successfully introduced and expressed in tuber-specific and constitutive manner. There was a striking increase in the growth and production of tubers in transgenic populations and also of the total protein content with an increase in most essential amino acids. The expressed protein was localized in the cytoplasm as well as in the vacuole of transgenic tubers. Thus we have been able to use a seed albumin gene with a well-balanced amino acid composition as a donor protein to develop a transgenic crop plant. The results document, in addition to successful nutritional improvement of potato tubers, the feasibility of genetically modifying other crop plants with novel seed protein composition.

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Section: Immunodetection Of Ama1 Protein In Transgenic Tuberssupporting

confidence: 92%

Increased nutritive value of transgenic potato by expressing a nonallergenic seed albumin gene from Amaranthus hypochondriacus

Chakraborty

Chakraborty²,

Datta³

2000

Proceedings of the National Academy of Sciences

131

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“…The CaMV 19s promoter directs the production of an abundant subgenomic mRNA encoding a protein that is both a major structural component of the cytosolic inclusion bodies and a translational activator for the polycistronic 35s RNA (Bonneville et al, 1989;Gowda et al, 1989). Although the CaMV 19s promoter is a strong promoter, it is reportedly 10-to 50-fold less active than the CaMV 35s promoter (Lawton et al, 1987). The CaMV 35s promoter directs the production of a terminally redundant transcript that is 180 nucleotides greater than genome length.…”

Section: Introductionmentioning

confidence: 99%

The Commelina yellow mottle virus promoter is a strong promoter in vascular and reproductive tissues.

1992

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Commelina yellow mottle virus (CoYMV) is a double-stranded DNA virus that infects the monocot Commelina diffusa. Although CoYMV and cauliflower mosaic virus (CaMV; another double-stranded DNA virus) probably replicate by a similar mechanism, the particle morphology and host range of CoYMV place it in a distinct group. We present evidence that a promoter fragment isolated from CoYMV confers a tissue-specific pattern of expression that is different from that conferred by the CaMV 35s promoter. When the CoYMV promoter is used to drive expression of the P-glucuronidase reporter gene in stably transformed tobacco plants, 0-glucuronidase activity occurs primarily in the phloem, the phloemassociated cells, and the axial parenchyma of roots, stems, leaves, and flowers. Activity is also detected throughout the anther, with highest activity in the tapetum. In contrast, the CaMV 35s promoter is active in most cell types. The CoYMV promoter is a strong promoter, and when the activity of the CoYMV promoter is compared with that of a duplicated CaMV 35s promoter, it is 30% as active in tobacco suspension cells and up to 25% as active in maize suspension cells. These properties of the CoYMV promoter make it potentially useful for high-leve1 expression of engineered genes in vascular cells.

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“…Although exceptions exist, several studies have shown that seed storage proteins are generally significantly less stable in the vegetative versus the seed tissues of a foreign host (Lawton et al 1987;Bagga et al 1992;Higgins and Spencer 1991;Nakamura et al 1993; but see also Utsumi et al 1993). For example, despite similar levels of 13-conglycinin mRNA in the leaves and seeds of transgenic petunia (utilizing a constitutive viral promoter to drive expression), the leaf cell environment yields a much lower level of protein accumulation (e.g., by 30-fold for the a'-subunit) (Lawton et al 1987;Nakamura et al 1993).…”

Section: Discussionmentioning

confidence: 94%

“…The low levels of accumulation of seed storage proteins observed in the transgenic host leaves, compared with the seed, even when the genes contain the cauliflower mosaic virus (CaMV) 35S promoter Lawton et al 1987;Bagga et al 1992;Nakamura et al 1993), may result from the continuous breakdown of the proteins in the protease-rich vacuoles of the leaves (Wandelt et al 1992). Differential proteolytic processing of storage proteins, yielding polypeptides of variable length in these two tissues, has also been documented, although the precise relationship between this and a lack of protein stability in the leaf, due to proteolysis, is presently unclear.…”

Section: Introductionmentioning

confidence: 98%

Accumulation and proteolytic processing of vicilin deletion-mutant proteins in the leaf and seed of transgenic tobacco

Kermode

Fisher

Polishchuk

et al. 1995

Planta

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Vicilin, a 7S globulin of Pisum sativum L. seed, accumulates in protein-storage vacuoles (protein bodies) of cotyledonary storage-parenchyma cells. The synthesis and proteolytic processing of various genetically engineered proteins within the leaf and seed of a heterologous (tobacco, Nicotiana tabacum L.) host was examined. A modified vicilin gene, in which the DNA sequence corresponding to the signal peptide was removed, resulted in a polypeptide of 50 kDa in the tobacco leaf and seed; none of the normal proteolytic cleavage products characteristic of expression of an unmodified vicilin gene were obtained. Likewise, no vacuolar accumulation of this mutant vicilin occurred in leaf protoplasts, which is also supportive of the predicted cytosolic localization for this protein. In-frame deletions were made within the region of the vicilin gene encoding the mature protein, to eliminate the N-terminal 28 and 121 amino acids and the C-terminal 69 residues, while maintaining an intact signal peptide. All of these "mature" deletion-mutant proteins were accumulated to only low levels in the host, but exhibited the predicted molecular weight and underwent some normal proteolytic processing in the seed. Mutant vicilin proteins having deletions in either the N-terminus (delta NT 121) or C-terminus (delta CT 69) were not found in appreciable amounts within the vacuolar fraction of transgenic tobacco leaf protoplasts, perhaps due to protein degradation in this compartment. Compared with the intact vicilin, oligomer assembly of the C-terminal deletion-mutant protein was disrupted in leaf cells, which may have further affected protein stability.(ABSTRACT TRUNCATED AT 250 WORDS)

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Expression of a soybean ?-conclycinin gene under the control of the Cauliflower Mosaic Virus 35S and 19S promoters in transformed petunia tissues

Cited by 60 publications

References 32 publications

Increased nutritive value of transgenic potato by expressing a nonallergenic seed albumin gene from Amaranthus hypochondriacus

Increased nutritive value of transgenic potato by expressing a nonallergenic seed albumin gene from Amaranthus hypochondriacus

The Commelina yellow mottle virus promoter is a strong promoter in vascular and reproductive tissues.

Accumulation and proteolytic processing of vicilin deletion-mutant proteins in the leaf and seed of transgenic tobacco

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