Cellulase gene expression in ripening avocado fruit: The accumulation of cellulase mRNA and protein as demonstrated by cDNA hybridization and immunodetection |
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Authors: | Rolf E Christoffersen Mark L Tucker George G Laties |
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Institution: | (1) Department of Biology and Molecular Biology Institute, University of California, 90024 LA, U.S.A.;(2) Present address: Mann Laboratory, University of California, 95616 Davis, CA, U.S.A.;(3) Present address: Department of Plant Molecular Biology, University of California, 94720 Berkeley, CA, U.S.A. |
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Abstract: | Summary A cDNA library was constructed from poly(A)+RNA of ripe avocado fruit. Colony hybridization identified a number of ripening specific clones of which one, pAV5, was shown to be specific for cellulase. Hybrid selection with pAV5 provided a message from ripe fruit that on in vitro translation yielded a polypeptide of 53kD, comigrating with purified avocado cellulase on SDS polyacrylamide gel electrophoresis. The translation product was selectively immunoprecipitated by antiserum to purified avocado cellulase. Immunoblotting of unripe and ripe avocado fruit extracts following SDS-PAGE showed a plentiful immunoreactive polypeptide in ripe fruit, and essentially none in unripe fruit. Hybridization of pAV5 to poly(A)+-RNA from unripe and ripe avocado fruit demonstrated that there is at least a 50-fold increase in the cellulase message concentration during ripening. Thus, the expression of cellulase enzyme activity during ripening is regulated by the appearance of mRNA coding for cellulase rather than by either translational or post-translational control mechanisms.Abbreviations poly(A)+
polyadenylated
- DS
sodium dodecyl sulfate
- D
kilodalton
- bp
base pairs
Supported by Research Grant GM 19807 from the United States Public Health Service and by additional funds from the University of California Research Council. |
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Keywords: | avocado cDNA-clone cellulase hybrid-select immunoblot poly(A)+RNA |
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