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Procedures for constructing ds-cDNA clone banks

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Plant Molecular Biology Manual

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Abstract

The construction of the first plant ds-cDNA clones used the abundant mRNAs which encode bean seed storage proteins of Phaseolus vulgaris and Glycine max as starting materials [11, 32, 33]. These mRNAs comprise about 30–40% of the total poly(A)+ RNA in developing cotyledons, and with just a few purification steps can be enriched to a purity of greater than 95% [33]. Construction and isolation of cDNA clones representing these bean storage protein mRNAs was relatively straightforward and did not require a high degree of efficiency in either construction, transformation, or screening procedures. In most cases, identification of the correct clone involved little more than checking the inserts from a few colonies by restriction enzyme digestion, followed by blot hybridization with 32P-labeled single-stranded (ss) cDNA [33].

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Author information

Authors and Affiliations

  1. Division of Molecular Biology, The Upjohn Company, Kalamazoo, MI, 49007, USA

    Jerry L. Slightom & Hector D. Quemada

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  1. Jerry L. Slightom
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  2. Hector D. Quemada
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Editors and Affiliations

  1. Purdue University, West Lafayette, Indiana, USA

    Stanton B. Gelvin

  2. Leiden State Universiy, Leiden, The Netherlands

    Robbert A. Schilperoort

  3. Ohio State University, Columbus, Ohio, USA

    Desh Pal S. Verma

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© 1989 Kluwer Academic Publishers, Dordrecht

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Slightom, J.L., Quemada, H.D. (1989). Procedures for constructing ds-cDNA clone banks. In: Gelvin, S.B., Schilperoort, R.A., Verma, D.P.S. (eds) Plant Molecular Biology Manual. Springer, Dordrecht. https://doi.org/10.1007/978-94-009-0951-9_7

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  • DOI: https://doi.org/10.1007/978-94-009-0951-9_7

  • Publisher Name: Springer, Dordrecht

  • Print ISBN: 978-94-010-6918-2

  • Online ISBN: 978-94-009-0951-9

  • eBook Packages: Springer Book Archive

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