Ma, H., Yanofsky, M. F., Klee, H. J., Bowman, J. L., Meyerowitz, E. M. (August 1992) Vectors for plant transformation and cosmid libraries. Gene, 117 (2). pp. 161-7. ISSN 0378-1119 (Print)0378-1119 (Linking)
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 (pCIT30), 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: | Paper |
|---|---|
| Uncontrolled Keywords: | Agrobacterium tumefaciens/*genetics Base Sequence Cloning, Molecular Cosmids/*genetics Gene Library Genetic Vectors/*genetics Molecular Sequence Data Plants/*genetics/microbiology Restriction Mapping Transformation, Genetic/*genetics |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing organism description > plant |
| CSHL Authors: | |
| Communities: | CSHL labs |
| Depositing User: | Matt Covey |
| Date: | 15 August 1992 |
| Date Deposited: | 01 Oct 2015 16:10 |
| Last Modified: | 01 Oct 2015 16:10 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/31793 |
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