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Design and development of amplifiable broad-host-range cloning vectors: analysis of the vir region of Agrobacterium tumefaciens plasmid pTiC58.
Plasmid 1984; 12(2):111-8P

Abstract

The construction of a set of new plasmids that are suitable as general cloning vectors in Escherichia coli and Agrobacterium tumefaciens is described. Plasmid pUCD2 is amplifiable in E. coli, replicates in a wide range of gram-negative hosts and contains a number of useful restriction endonuclear cleavage sites and antibiotic resistance genes. This includes unique sites for KpnI, SacI, SacII, PstI, ClaI, SalI, EcoRV, and PvuII and the genes for resistance to kanamycin, tetracycline, ampicillin, and spectinomycin/streptomycin. Derivatives of pUCD2 include pUCD4, which has a unique XbaI site and the cosmid pUCD5, which also contains a unique EcoRI site. Two smaller plasmids pUCD9P and pUCD9X, contain many of the same unique sites as pUCD2 and pUCD4, but carry only the pBR322 replication origin and therefore do not display the extensive host-range of pSa. These plasmids were used to isolate and manipulate fragments of the A. tumefaciens pTiC58 plasmid in both E. coli and A. tumefaciens. Fragments from the virulence (vir) region of pTiC58 inserted immediately upstream of the spectinomycin resistance gene of pUCD2 resulted in spectinomycin resistance levels that varied greatly depending on the particular fragment and its orientation of insertion. Using this property we find that a major portion of the vir region of pTiC58 is transcribed in A. tumefaciens and E. coli from left to right toward the T region.

Authors

No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

6095350

Citation

Close, T J., et al. "Design and Development of Amplifiable Broad-host-range Cloning Vectors: Analysis of the Vir Region of Agrobacterium Tumefaciens Plasmid PTiC58." Plasmid, vol. 12, no. 2, 1984, pp. 111-8.
Close TJ, Zaitlin D, Kado CI. Design and development of amplifiable broad-host-range cloning vectors: analysis of the vir region of Agrobacterium tumefaciens plasmid pTiC58. Plasmid. 1984;12(2):111-8.
Close, T. J., Zaitlin, D., & Kado, C. I. (1984). Design and development of amplifiable broad-host-range cloning vectors: analysis of the vir region of Agrobacterium tumefaciens plasmid pTiC58. Plasmid, 12(2), pp. 111-8.
Close TJ, Zaitlin D, Kado CI. Design and Development of Amplifiable Broad-host-range Cloning Vectors: Analysis of the Vir Region of Agrobacterium Tumefaciens Plasmid PTiC58. Plasmid. 1984;12(2):111-8. PubMed PMID: 6095350.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Design and development of amplifiable broad-host-range cloning vectors: analysis of the vir region of Agrobacterium tumefaciens plasmid pTiC58. AU - Close,T J, AU - Zaitlin,D, AU - Kado,C I, PY - 1984/9/1/pubmed PY - 1984/9/1/medline PY - 1984/9/1/entrez SP - 111 EP - 8 JF - Plasmid JO - Plasmid VL - 12 IS - 2 N2 - The construction of a set of new plasmids that are suitable as general cloning vectors in Escherichia coli and Agrobacterium tumefaciens is described. Plasmid pUCD2 is amplifiable in E. coli, replicates in a wide range of gram-negative hosts and contains a number of useful restriction endonuclear cleavage sites and antibiotic resistance genes. This includes unique sites for KpnI, SacI, SacII, PstI, ClaI, SalI, EcoRV, and PvuII and the genes for resistance to kanamycin, tetracycline, ampicillin, and spectinomycin/streptomycin. Derivatives of pUCD2 include pUCD4, which has a unique XbaI site and the cosmid pUCD5, which also contains a unique EcoRI site. Two smaller plasmids pUCD9P and pUCD9X, contain many of the same unique sites as pUCD2 and pUCD4, but carry only the pBR322 replication origin and therefore do not display the extensive host-range of pSa. These plasmids were used to isolate and manipulate fragments of the A. tumefaciens pTiC58 plasmid in both E. coli and A. tumefaciens. Fragments from the virulence (vir) region of pTiC58 inserted immediately upstream of the spectinomycin resistance gene of pUCD2 resulted in spectinomycin resistance levels that varied greatly depending on the particular fragment and its orientation of insertion. Using this property we find that a major portion of the vir region of pTiC58 is transcribed in A. tumefaciens and E. coli from left to right toward the T region. SN - 0147-619X UR - https://www.unboundmedicine.com/medline/citation/6095350/Design_and_development_of_amplifiable_broad_host_range_cloning_vectors:_analysis_of_the_vir_region_of_Agrobacterium_tumefaciens_plasmid_pTiC58_ L2 - https://linkinghub.elsevier.com/retrieve/pii/0147-619X(84)90057-X DB - PRIME DP - Unbound Medicine ER -