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Tobacco cytochromeb 5: cDNA isolation, expression analysis andin vitro protein targeting

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Abstract

A full-length clone encoding cytochromeb 5 has been isolated from a tobacco leaf cDNA library in αgt11 by PCR using degenerate primers. This cDNA encodes a protein of 139 residues which exhibits a high degree of homology to other cytochromeb 5s, the message for which is expressed predominantly in developing seeds and in pigmented flower tissue. In the developing tobacco seed the mRNA is abundant at very early stages (<10 days after flowering). Southern analysis indicated that more than one gene encodes cytochromeb 5 in the tobacco genome.In vitro transcription and translation studies of the cDNA indicated that the protein inserts into the ER membrane by a non-SRP-mediated pathway and that theC-terminus of the protein is required for targeting and insertion.

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Authors and Affiliations

  1. Department of Botany, University of Bristol, BS8 1UG, Bristol, UK

    Mark A. Smith & A. Keith Stobart

  2. Department of Agricultural Sciences, University of Bristol, Institute of Arable Crops Research, Long Ashton Research Station, Bristol, BS18 9AF, UK

    Peter R. Shewry & Johnathan A. Napier

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  1. Mark A. Smith
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  2. A. Keith Stobart
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  3. Peter R. Shewry
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  4. Johnathan A. Napier
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Smith, M.A., Stobart, A.K., Shewry, P.R. et al. Tobacco cytochromeb 5: cDNA isolation, expression analysis andin vitro protein targeting. Plant Mol Biol 25, 527–537 (1994). https://doi.org/10.1007/BF00043880

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  • DOI: https://doi.org/10.1007/BF00043880

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