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J Tempé

Publications and source records attributed to J Tempé.

28 records · Page 2Linked to original sources

Structure of T-DNA in roots transformed by Agrobacterium rhizogenes.

DNA isolated from hairy roots incited on Daucus carota by Agrobacterium strains harboring the Ri plasmid was probed with Ri plasmid fragments to identify plasmid sequences transferred and integrated into the plant genome. One hairy root line was incited by an A. rhizogenes natural isolate strain harboring two large plasmids in addition to the Ri plasmid. DNA from hairy roots incited by this strain contained a total of five nonidentical copies of T-DNA which together comprised a T-DNA complement representing a 34-42 kb segment of the Ri plasmid. A second hairy root line was incited by a Ti plasmidless A. tumefaciens strain into which the Ri plasmid had been conjugated. Hairy root DNA incited by this strain contained four identical T-DNA copies 17-18 kb in length and one truncated copy. T-DNA borders occurred at preferential sites of the Ri plasmid. T-DNA structure in these hairy root lines is similar to T-DNA structure in tumors incited by Ti plasmids of A. tumefaciens.

DNA↗

Agropine in "null-type" crown gall tumors: Evidence for generality of the opine concept.

Agrobacterium Ti (tumor-inducing) plasmids, the causative agents of crown gall disease, fall into four genetic groups based on the patterns of octopine and nopaline synthesis (by crown gall tumors) and catabolism (by Agrobacterium tumefaciens) for which they are responsible. Two classes of Ti plasmids induce tumors that synthesize neither octopine nor nopaline. The existence of these Ti plasmids challenged the view that opines such as octopine and nopaline play a central role in crown gall biology. We now report the occurrence of an opine in tumors induced by one of these classes of Ti plasmids, the "null-type" plasmids typified by pTi Bo542. The opine was purified by biological enrichment based on its utilization by bacteria containing pTi Bo542 but not by bacteria lacking a Ti plasmid. The mass spectrum and biological properties of this opine are identical to those of agropine, an opine recently discovered in octopine-type tumors. We propose that null-type Ti plasmids now be named for their signal opine, agropine-type Ti plasmids.

Journal Article↗

Thermosensitive step associated with transfer of the Ti plasmid during conjugation: Possible relation to transformation in crown gall.

It is reported here that transfer by means of a conjugative process of an oncogenic plasmid from a virulent strain of Agrobacterium tumefaciens to a strain of that organism that had been cured of the plasmid is thermosensitive. Since the thermosensitive step found in the conjugative process appears similar in every respect to a thermosensitive step that is involved in the transformation of a normal cell to a tumor cell in the crown gall disease of plants, it is suggested that the observed results may reflect the existence of a thermosensitive step that is common to these two processes.

Journal Article↗

Localization and characterization of the region encoding catabolism of mannopinic acid from the octopine-type Ti plasmid pTi15955.

The region of the octopine-type tumor-inducing (Ti) plasmid pTi15955 encoding catabolism of mannopinic acid was localized to an 18-kilobase (kb) segment mapping between Ti plasmid coordinates 70 and 88. A subclone containing only this region normally did not allow utilization of any other mannityl opine. However, spontaneous mutants of the plasmid were isolated that conferred catabolism of agropinic acid. While the mutants remained regulated for mannopinic acid utilization, induction by this opine resulted in increased activity associated with agropinic acid catabolism. Respirometric studies and results from growth assays on mannopinic acid analogues indicated that the genes encoded on the 18-kb subclone were regulated in a wild-type fashion. By means of these analogues, spontaneous constitutive mutants were isolated. In several cases, the mutant phenotype was associated with an insertion event mapping within a 300-base pair region of the subclone insert. Although constitutive for catabolism of mannopinic acid, these mutants remained unable to catabolize any of the other mannityl opines. Segregation studies and genetic reconstitution experiments showed that one of the subclones isolated directly in Agrobacterium was actually composed of two complementing recombinant plasmids contained in the same cell. This indicated that the genes encoding mannopinic acid catabolism were organized into at least two complementation groups. These results point to a high degree of complexity in the organization and regulation of Ti plasmid genes encoding catabolism of a relatively simple carbon source.

DNA Restriction Enzymes↗