scholarly journals Review: Plant Binary Vectors of Ti Plasmid in <i>Agrobacterium tumefaciens</i> with a Broad Host-Range Replicon of pRK2, pRi, pSa or pVS1

2013 ◽  
Vol 04 (04) ◽  
pp. 932-939 ◽  
Author(s):  
Norimoto Murai
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Host Range ◽  
Ti Plasmid ◽  
Broad Host Range ◽  
Binary Vectors

scholarly journals Construction of a Derivative of Agrobacterium tumefaciens C58 That Does Not Mutate to Tetracycline Resistance

2001 ◽  
Vol 14 (1) ◽  
pp. 98-103 ◽  
Author(s):  
Zhao-Qing Luo ◽  
Thomas E. Clemente ◽  
Stephen K. Farrand
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Host Range ◽  
High Frequency ◽  
Tetracycline Resistance ◽  
Physiological Traits ◽  
Selection Marker ◽  
Detectable Effect ◽  
Broad Host Range ◽  
Binary Vectors ◽  
Agrobacterium Tumefaciens C58

Agrobacterium tumefaciens C58 mutates to tetracycline resistance at high frequency, complicating the use of many broad-host-range cloning and binary vectors that code for resistance to this antibiotic as the selection marker. Such mutations are associated with a resistant gene unit, tetC58, that is present in the genome of this strain. By deleting the tetC58 locus, we constructed NTL4, a derivative of C58 that no longer mutates to tetracycline resistance. The deletion had no detectable effect on genetic or physiological traits of NTL4 or on the ability of this strain to transform plants.

Host range of Agrobacterium tumefaciens is determined by the Ti plasmid

Nature ◽  
1980 ◽  
Vol 283 (5749) ◽  
pp. 794-796 ◽  
Author(s):  
M. F. Thomashow ◽  
C. G. Panagopoulos ◽  
M. P. Gordon ◽  
E. W. Nester
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Host Range ◽  
Ti Plasmid

scholarly journals Broad-Host-Range Expression Vectors with Tightly Regulated Promoters and Their Use To Examine the Influence of TraR and TraM Expression on Ti Plasmid Quorum Sensing

2008 ◽  
Vol 74 (16) ◽  
pp. 5053-5062 ◽  
Author(s):  
Sharik R. Khan ◽  
Jennifer Gaines ◽  
R. Martin Roop ◽  
Stephen K. Farrand
Keyword(s):  
Host Range ◽  
Gene Activation ◽  
Ti Plasmid ◽  
Expression Vectors ◽  
Multiple Cloning Site ◽  
Broad Host Range ◽  
Precise Control ◽  
Broad Host Range Plasmid ◽  
Cloned Gene ◽  
Ndei Site

ABSTRACT Experiments requiring strong repression and precise control of cloned genes can be difficult to conduct because of the relatively high basal level of expression of currently employed promoters. We report the construction of a family of vectors that contain a reengineered lacI q-lac promoter-operator complex in which cloned genes are strongly repressed in the absence of inducer. The vectors, all based on the broad-host-range plasmid pBBR1, are mobilizable and stably replicate at moderate copy number in representatives of the alpha- and gammaproteobacteria. Each vector contains a versatile multiple cloning site that includes an NdeI site allowing fusion of the cloned gene to the initiation codon of lacZα. In each tested bacterium, a uidA reporter fused to the promoter was not expressed at a detectable level in the absence of induction but was inducible by 10- to 100-fold, depending on the bacterium. The degree of induction was controllable by varying the concentration of inducer. When the vector was tested in Agrobacterium tumefaciens, a cloned copy of the traR gene, the product of which is needed at only a few copies per cell, did not confer activity under noninducing conditions. We used this attribute of very tight and variably regulatable control to assess the relative amounts of TraR required to activate the Ti plasmid conjugative transfer system. We identified levels of induction that gave wild-type transfer frequencies, as well as levels that induced correspondingly lower frequencies of transfer. We also used this system to show that the antiactivator TraM sets the level of intracellular TraR required for tra gene activation.

scholarly journals A T-DNA from the Agrobacterium tumefaciens Limited-Host-Range Strain AB2/73 Contains a Single Oncogene

1998 ◽  
Vol 11 (5) ◽  
pp. 335-342 ◽  
Author(s):  
Léon Otten ◽  
Julien Schmidt
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Host Range ◽  
Dna Sequence ◽  
Ti Plasmid ◽  
Dna Homology ◽  
Agrobacterium Vitis ◽  
E Gene ◽  
Limited Host Range ◽  
Border Sequences ◽  
Border Sequence

Agrobacterium tumefaciens strain AB2/73 isolated from Lippia canescens has been described as a limited-host-range strain. Its tumor-inducing (Ti) plasmid has been found to lack DNA homology to known T-DNAs (L. Unger, S. F. Ziegler, G. A. Huffman, V. C. Knauf, R. Peet, L. W. Moore, M. P. Gordon, and E. W. Nester. J. Bacteriol. 164:723–730, 1985). We have isolated a T-DNA from AB2/73 by using a heterologous border sequence as a probe. The AB2/73 T-DNA sequence (3,504 bp) is flanked by canonical border sequences, has no detectable DNA homology with other T-DNAs, and contains only two genes: lsn ( Lippia strain nopaline synthaselike gene) and lso ( Lippia strain oncogene). The lso gene induces nondif-ferentiating tumors on a limited number of hosts when transferred by a Ti plasmid from a wide-host-range strain. Part of the predicted Lso protein is weakly homologous to other Agrobacterium oncoproteins encoded by rolB, rolBTR, orf13, gene e, gene 5, and gene 3′. A 28-kb fragment corresponding to the virA to virE region was cloned by using a heterologous vir fragment as probe. The AB2/73 vir region is homologous to most of the C58 virulence region; however, the virA gene is most related to the virA gene of the Agrobacterium vitis limited-host-range strain Ag162.

scholarly journals Construction of Disarmed Ti Plasmids Transferable between Escherichia coli and Agrobacterium Species

2009 ◽  
Vol 75 (7) ◽  
pp. 1845-1851 ◽  
Author(s):  
Kazuya Kiyokawa ◽  
Shinji Yamamoto ◽  
Kei Sakuma ◽  
Katsuyuki Tanaka ◽  
Kazuki Moriguchi ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Host Range ◽  
Plant Transformation ◽  
Ti Plasmid ◽  
Broad Host Range ◽  
Bacterial Strains ◽  
E Coli ◽  
Ti Plasmids ◽  
The Right ◽  
Border Sequence

ABSTRACT Agrobacterium-mediated plant transformation has been used widely, but there are plants that are recalcitrant to this type of transformation. This transformation method uses bacterial strains harboring a modified tumor-inducing (Ti) plasmid that lacks the transfer DNA (T-DNA) region (disarmed Ti plasmid). It is desirable to develop strains that can broaden the host range. A large number of Agrobacterium strains have not been tested yet to determine whether they can be used in transformation. In order to improve the disarming method and to obtain strains disarmed and ready for the plant transformation test, we developed a simple scheme to make certain Ti plasmids disarmed and simultaneously maintainable in Escherichia coli and mobilizable between E. coli and Agrobacterium. To establish the scheme in nopaline-type Ti plasmids, a neighboring segment to the left of the left border sequence, a neighboring segment to the right of the right border sequence of pTi-SAKURA, a cassette harboring the pSC101 replication gene between these two segments, the broad-host-range IncP-type oriT, and the gentamicin resistance gene were inserted into a suicide-type sacB-containing vector. Replacement of T-DNA with the cassette in pTiC58 and pTi-SAKURA occurred at a high frequency and with high accuracy when the tool plasmid was used. We confirmed that there was stable maintenance of the modified Ti plasmids in E. coli strain S17-1λpir and conjugal transfer from E. coli to Ti-less Agrobacterium strains and that the reconstituted Agrobacterium strains were competent to transfer DNA into plant cells. As the modified plasmid delivery system was simple and efficient, conversion of strains to the disarmed type was easy and should be applicable in studies to screen for useful strains.

scholarly journals Identification of Rhizobium plasmid sequences involved in recognition of Psophocarpus, Vigna, and other legumes.

1986 ◽  
Vol 102 (4) ◽  
pp. 1173-1182 ◽  
Author(s):  
W J Broughton ◽  
C H Wong ◽  
A Lewin ◽  
U Samrey ◽  
H Myint ◽  
...  
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Host Range ◽  
Dna Sequences ◽  
Nodule Development ◽  
Broad Host Range ◽  
Psophocarpus Tetragonolobus ◽  
Rhizobium Species ◽  
Symbiotic Plasmid ◽  
Slow Growing ◽  
Overlapping Sets

Symbiotic DNA sequences involved in nodulation by Rhizobium must include genes responsible for recognizing homologous hosts. We sought these genes by mobilizing the symbiotic plasmid of a broad host-range Rhizobium MPIK3030 (= NGR234) that can nodulate Glycine max, Psophocarpus tetragonolobus, Vigna unguiculata, etc., into two Nod- Rhizobium mutants as well as into Agrobacterium tumefaciens. Subsequently, cosmid clones of pMPIK3030a were mobilized into Nod+ Rhizobium that cannot nodulate the chosen hosts. Nodule development was monitored by examining the ultrastructure of nodules formed by the transconjugants. pMPIK3030a could complement Nod- and Nif- deletions in R. leguminosarum and R. meliloti as well as enable A. tumefaciens to nodulate. Three non-overlapping sets of cosmids were found that conferred upon a slow-growing Rhizobium species, as well as on R. loti and R. meliloti, the ability to nodulate Psophocarpus and Vigna, thus pointing to the existence of three sets of host-specificity genes. Recipients harboring these hsn regions had truly broadened host-range since they could nodulate both their original hosts as well as MPIK3030 hosts.

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