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Vectors for plant transformation and cosmid libraries

Ma, Hong and Yanofsky, Martin F. and Klee, Harry J. and Bowman, John L. and Meyerowitz, Elliot M. (1992) Vectors for plant transformation and cosmid libraries. Gene, 117 (2). pp. 161-167. ISSN 0378-1119. https://resolver.caltech.edu/CaltechAUTHORS:20161214-165342441

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Abstract

A series of vectors has been constructed for the purpose of introducing cloned DNAs into plant genomes, using Agrobacterium tumefaciens-mediated transformation methods. One of these vectors, pCIT20, is a plasmid that contains a multiple cloning site (MCS), and a marker (Hph) that confers hygromycin resistance to plant cells. The others are all cosmid vectors which allow insertion of up to 46 kb of plant genomic DNA, and which also contain all of the necessary sequences for A. tumefaciens-mediated plant transformation. The cosmid vectors either contain a Hph marker (pCIT03), or a kanamycin-resistance marker (pCIT101–104). Three of the cosmid vectors (pCIT30, pCIT101, and pCIT103) carry bacteriophage T7 and SP6 promoters flanking the cloning Bg/II site, for synthesis of end-specific RNAs. The end-specific RNAs may be used as probes when labeled with radioactive or biotinylated nucleotides, for example, in a chromosome-walking experiment. The other two cosmid vectors (pCIT102 and pCIT104) carry restriction sites flanking the insertion site (XhoI) for convenient release of the insert by restriction digests. These sites, in combination with sites internal to the insert, allow the generation of end fragments for subcloning or labeling probes. These vectors should be valuable for isolation and analysis of plant genes, using transformation, library screening, and chromosome-walking approaches.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1016/0378-1119(92)90725-5DOIArticle
http://www.sciencedirect.com/science/article/pii/0378111992907255PublisherArticle
ORCID:
AuthorORCID
Meyerowitz, Elliot M.0000-0003-4798-5153
Additional Information:© 1992 Elsevier. Received 18 December 1991, Revised 5 March 1992, Accepted 10 March 1992. We thank H. Huang and A. Tagle for assistance. We would also like to thank Alan Lloyd and Yuonne Muskopf for their help in constructing pMON754 and pMON721. This work was supported by grants from the National Science Foundation (DCB 8703439) and from the Lucille P. Markey Charitable Trust to E.M.M., and by grants from the National Science Foundation (DCB 9004567 and DCB 9105260) and funds from the Robertson Foundation to H.M.H.M. was a Helen Hay Whitney post-doctoral fellow, M.F.Y. was a National Science Foundation plant biology post-doctoral fellow, and J.L.B. was supported by a National Institutes of Health pre-doctoral training grant.
Funders:
Funding AgencyGrant Number
NSFDCB-8703439
Lucille P. Markey Charitable TrustUNSPECIFIED
NSFDCB-9004567
NSFDCB-9105260
Robertson FoundationUNSPECIFIED
Helen Hay Whitney FoundationUNSPECIFIED
NSF Plant Biology Postdoctoral FellowshipUNSPECIFIED
NIH Predoctoral FellowshipUNSPECIFIED
Subject Keywords:Agrobacterium; binary vector; chromosome walking; end-specific probe; hygromycin; integrative vector; plasmid; recombinant DNA
Issue or Number:2
Record Number:CaltechAUTHORS:20161214-165342441
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20161214-165342441
Official Citation:Hong Ma, Martin F. Yanofsky, Harry J. Klee, John L. Bowman, Elliot M. Meyerowitz, Vectors for plant transformation and cosmid libraries, Gene, Volume 117, Issue 2, 15 August 1992, Pages 161-167, ISSN 0378-1119, http://dx.doi.org/10.1016/0378-1119(92)90725-5. (http://www.sciencedirect.com/science/article/pii/0378111992907255)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:72841
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:15 Dec 2016 18:18
Last Modified:03 Oct 2019 16:21

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