Regulation of an Arabidopsis oleosin gene promoter in transgenic Brassica napus

Plant, Áine L.; Rooijen, Gijs J. H. van; Anderson, Clarke P.; Moloney, Maurice M.

doi:10.1007/bf00023237

Cited by 39 publications

(21 citation statements)

References 59 publications

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“…In Figure 7, it can be seen that wild-type B. carinata produced no detectable signals, whereas B. napus GB5 plants (oleosin promoter-GUS; Plant et al, 1994) gave a strong reaction to GUS antibodies at the expected size of free GUS (68 kD). OBPGUSA generated the expected full-length product at 86 kD.…”

Section: Proline Knot Motif Is Critical In Vivo For Oil Body Targetinmentioning

confidence: 99%

Role of the Proline Knot Motif in Oleosin Endoplasmic Reticulum Topology and Oil Body Targeting

Abell¹,

Holbrook²,

Abenes³

et al. 1997

The Plant Cell

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105

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An Arabidopsis oleosin was used as a model to study oleosin topology and targeting to oil bodies. Oleosin mRNA was in vitro translated with canine microsomes in a range of truncated forms. This allowed proteinase K mapping of the membrane topology. Oleosin maintains a conformation with a membrane-integrated hydrophobic domain flanked by N-and C-terminal domains located on the outer microsome surface. This is a unique membrane topology on the endoplasmic reticulum (ER). Three universally conserved proline residues within the "proline knot" motif of the oleosin hydrophobic domain were substituted by leucine residues. After in vitro translation, only minor differences in proteinase K protection could be observed. These differences were not apparent in soybean microsomes. No significant difference in incorporation efficiency on the ER was observed between the two oleosin forms. However, as an oleosin-P-glucuronidase translational fusion, the proline knot variant failed to target to oil bodies in both transient embryo expression and in stably transformed seeds. Fractionation of transgenic embryos expressing oleosin-P-glucuronidase fusions showed that the proline knot variant accumulated in the ER to similar levels compared with the native form. Therefore, the proline knot motif is not important for ER integration and the determination of topology but is required for oil body targeting. The loss of the proline knot results in an intrinsic instability in the oleosin polypeptide during trafficking.

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Section: Proline Knot Motif Is Critical In Vivo For Oil Body Targetinmentioning

confidence: 99%

Role of the Proline Knot Motif in Oleosin Endoplasmic Reticulum Topology and Oil Body Targeting

Abell¹,

Holbrook²,

Abenes³

et al. 1997

The Plant Cell

Self Cite

105

View full text Add to dashboard Cite

show abstract

“…Using an oleosin promoter from protein thioesterase from seeds of the California Bay {Umbel-Arabidopsis fused to the gus gene. Plant et al (1994) performed lularia californica) resulting in plants accumulating up to 50% truncation-and activation analysis of oilseed rape. The GUS lauric acid (Voelker et al 1992).…”

Section: Assessment Of Transgenic Plantsmentioning

confidence: 99%

“…Applying an oleosin gene promoter from Arabidopsis, the ^-glucoronidase gene product was targeted to the oleosomes of Brassica napus (Plant et al 1994). This property can be exploited for the production and partial purification of recombinant proteins or directly applied as oilbody-immobilized enzymes in industrial processes (van Rooijen and Moloney 1995).…”

Section: Other Traitsmentioning

confidence: 99%

Genetic transformation of Brassica

Poulsen

1996

Plant Breeding

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Tissue culture regeneration of plants from Brassica species can be achieved from various explant types. Their performance, however, varies with the genotype and age of the explant and is sensitive to medium supplements and Agrobacterium cocultivation. Most Brassica transformations were carried out using Agrobacterium and it was established that nopaline and agropine strains of .4. tumefaciens and A. rhizogenes, respectively, are the most efficient. Application of virulence gene induction is ambiguous. Binary vectors are mostly apphed in spite of indications that cointegrate vectors are more efficient. Cotransformation is successful with two different bacteria or with one bacterium carrying both plasmids, however, it has proved problematic to integrate the two transgenes into separate loci. Transformations with wild type rol genes develop hairy roots, from which transgenic plants can be regenerated. Vectorless transformation is feasible with microprojectile bombardment of microspore derived embryos being the most promising.Antibiotic markers providing resistance to kanamycin, hygromycin, and spectinomycin are applicable as selectable markers, while chloramphenicol is not suitable. Among herbicides, phosphinotricin gives good results while chlorsulfuron is unsuitable for in vitro selection. Major therapeutic antibiotics being successfully applied are carbenicillin, cefotaxime, amoxycillin, and ticarcillin. Standard methods for evaluating transgenic plants also have been applied to Brassica species. Some breeding objectives obtained through genetic transformation are presented.

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“…Abscisic acid (ABA) also plays an important role in expression of seed-storage protein genes and embryogenesis related genes (Luo et al, 2008). Plant et al (1994) showed that ABA positively regulated expression of GUS in transgenic Brassica containing the oleosin promoter from Arabidopsis fused to the reporter gene. Rowley and Herman (Rowley and Herman, 1997) found that soybean oleosin genes contain regulatory elements in the upstream region which also were found in vacuolar storage protein genes.…”

Section: Discussionmentioning

confidence: 99%

Reporter Gene Expression Patterns Regulated by an Ara h 2 Promoter Differ in Homologous Versus Heterologous Systems1

Bhattacharya¹,

Ramos²,

Faustinelli

et al. 2012

Peanut Science

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Peanut (Arachis hypogaea L.) is a globally important crop whose seeds are widely used in food products. Peanut seeds contain proteins that serve a nutrient reservoir function and that also are major allergens. As part of an investigation to determine the effect of reducing/ eliminating the peanut allergen Ara h 2 from seeds, gene sequence including upstream regulatory regions was characterized. The ability of regions upstream of the translation initiation site to regulate seed-specific expression of reporter genes was tested in peanut and Arabidopsis. Two independent transgenic peanut lines biolistically transformed with 1kb of DNA upstream of the Ara h 2.02 (B-genome) coding sequence controlling a Green Fluorescent Protein -b-glucuronidase (Gfp-Gus) fusion were obtained. All T 1 , T 2 and T 3 generations of transgenic plants showed the expression of GFP and GUS restricted to seeds and near background levels in vegetative tissues. However, constitutive GUS expression was observed in Arabidopsis transgenic lines, a heterologous system. It is possible that transacting factors regulating seed specificity in peanut are too divergent in Arabidopsis to enable the seed specific response. Thus, the promoter described in this paper may have potential use for expression of transgenes in peanut where seed-specificity is desired, but expression patterns should be tested in heterologous systems prior to off-the-shelf adoption.

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Regulation of an Arabidopsis oleosin gene promoter in transgenic Brassica napus

Cited by 39 publications

References 59 publications

Role of the Proline Knot Motif in Oleosin Endoplasmic Reticulum Topology and Oil Body Targeting

Role of the Proline Knot Motif in Oleosin Endoplasmic Reticulum Topology and Oil Body Targeting

Genetic transformation of Brassica

Reporter Gene Expression Patterns Regulated by an Ara h 2 Promoter Differ in Homologous Versus Heterologous Systems1

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