Section 8
Chapter 7,809

Specialized binary vector for plant transformation: expression of the Arabidopsis thaliana AHAS gene in Nicotiana tabacum

Olszewski, N.E.; Martin, F.B.; Ausubel, F.M.

Nucleic Acids Research 16(22): 10765-10782


ISSN/ISBN: 0305-1048
PMID: 3060849
DOI: 10.1093/nar/16.22.10765
Accession: 007808813

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We constructed a cosmid vector, pOCA18, designed for transferring plant genomic libraries from Agrobacterium tumefaciens to plants. Clones from a genomic library of Arabidopsis thaliana DNA in pOCA18 were propagated stably in both Escherichia coli and A. tumefaciens. Clones from the pOCA 18 A. thaliana library were used to construct transgenic Nicotiana tabacum plants; the DNA inserts were transferred intact in 10 out of 16 transgenic N. tabacum plants examined but were partially deleted in six others. Transgenic N. tabacum plants constructed with a mutant A. thaliana acetohydroxy acid synthase gene (from the pOCA18 library) that encodes an enzyme resistant to the herbicide chlorsulfuron were resistant to chlorsulfuron. A statistical analysis indicated that if the A. thaliana library contains 107 members and if 107 A. tumefaciens transconjugants containing the library were used to transform plants cells, then 2 .times. 104 transformed plant cells must be generated to have a 95% probability of constructing a transgenic plant carrying a specific DNA sequence from the A. thaliana library.

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