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J Bacteriol. Mar 1989; 171(3): 1665–1672.
PMCID: PMC209796

Sequence determination and characterization of the replicator region in the tumor-inducing plasmid pTiB6S3.


The replicator region of the 195-kilobase-pair (kb) tumor-inducing plasmid pTiB6S3 was previously identified by isolation of a 6.8-kb miniplasmid (B.P. Koekman, P.J.J. Hooykaas, and R.A. Schilperoort, Plasmid 7:119-132, 1982). This miniplasmid was joined to ColE1-based vectors and subjected to mutagenesis. The resulting mutant plasmids were examined for their ability to replicate autonomously in Agrobacterium tumefaciens. It was found that a 4.2-kb region was sufficient for displaying replication characteristics similar to those of the parental pTiB6S3. Nucleotide sequence analysis of this 4.2-kb region revealed the presence of three possible reading frames in the same direction (repA, repB, and repC). Proteins coded for by these frames were identified by in vitro synthesis in a coupled transcription-translation system. The replicating ability became attenuated by repA and repB mutations but was completely abolished by repC mutations. The size, arrangement, and mutational effects of the three rep genes were quite similar to those of the rep genes that were previously identified in the hairy root-inducing plasmid pRiA4b. However, defects caused by rep mutations in one plasmid were unable to be complemented by corresponding functions in the other plasmid.

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  • An G, Watson BD, Stachel S, Gordon MP, Nester EW. New cloning vehicles for transformation of higher plants. EMBO J. 1985 Feb;4(2):277–284. [PMC free article] [PubMed]
  • Costantino P, Hooykaas PJ, den Dulk-Ras H, Schilperoort RA. Tumor formation and rhizogenicity of Agrobacterium rhizogenes carrying Ti plasmids. Gene. 1980 Oct;11(1-2):79–87. [PubMed]
  • Das A, Stachel S, Ebert P, Allenza P, Montoya A, Nester E. Promoters of Agrobacterium tumefaciens Ti-plasmid virulence genes. Nucleic Acids Res. 1986 Feb 11;14(3):1355–1364. [PMC free article] [PubMed]
  • Furth ME, Dove WF, Meyer BJ. Specificity determinants for bacteriophage lambda DNA replication. III. Activation of replication in lambda ric mutants by transcription outside of ori. J Mol Biol. 1982 Jan 5;154(1):65–83. [PubMed]
  • Hirayama T, Muranaka T, Ohkawa H, Oka A. Organization and characterization of the virCD genes from Agrobacterium rhizogenes. Mol Gen Genet. 1988 Aug;213(2-3):229–237. [PubMed]
  • Holsters M, Silva B, Van Vliet F, Genetello C, De Block M, Dhaese P, Depicker A, Inzé D, Engler G, Villarroel R, et al. The functional organization of the nopaline A. tumefaciens plasmid pTiC58. Plasmid. 1980 Mar;3(2):212–230. [PubMed]
  • Hooykaas PJ, Hofker M, den Dulk-Ras H, Schilperoort RA. A comparison of virulence determinants in an octopine Ti plasmid, a nopaline Ti plasmid, and an Ri plasmid by complementation analysis of Agrobacterium tumefaciens mutants. Plasmid. 1984 May;11(3):195–205. [PubMed]
  • Hooykaas PJ, Schilperoort RA. The molecular genetics of crown gall tumorigenesis. Adv Genet. 1984;22:209–283. [PubMed]
  • Koekman BP, Hooykaas PJ, Schilperoort RA. Localization of the replication control region on the physical map of the octopine Ti plasmid. Plasmid. 1980 Sep;4(2):184–195. [PubMed]
  • Koekman BP, Hooykaas PJ, Schilperoort RA. A functional map of the replicator region of the octopine Ti plasmid. Plasmid. 1982 Mar;7(2):119–132. [PubMed]
  • Komari T, Halperin W, Nester EW. Physical and functional map of supervirulent Agrobacterium tumefaciens tumor-inducing plasmid pTiBo542. J Bacteriol. 1986 Apr;166(1):88–94. [PMC free article] [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Lark KG. Evidence for the direct involvement of RNA in the initiation of DNA replication in Escherichia coli 15T. J Mol Biol. 1972 Feb 28;64(1):47–60. [PubMed]
  • Sanger F. Determination of nucleotide sequences in DNA. Science. 1981 Dec 11;214(4526):1205–1210. [PubMed]
  • White FF, Nester EW. Relationship of plasmids responsible for hairy root and crown gall tumorigenicity. J Bacteriol. 1980 Nov;144(2):710–720. [PMC free article] [PubMed]
  • Yanisch-Perron C, Vieira J, Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. [PubMed]
  • Zubay G. In vitro synthesis of protein in microbial systems. Annu Rev Genet. 1973;7:267–287. [PubMed]

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