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. 1980 May;65(5):834–837. doi: 10.1104/pp.65.5.834

Partial Purification and Characterization of the mRNA for α-Amylase from Barley Aleurone Layers 1

Thomas J Mozer 1,2
PMCID: PMC440433  PMID: 16661291

Abstract

The poly(A)-containing mRNA from barley aleurone layers pretreated with gibberellic acid has been purified by phenol-chloroform extraction and repeated oligo[d(pT)]-cellulose chromatography. This RNA has been translated in both the wheat germ and reticulocyte lysate in vitro translation systems with greater than 50% of the synthesized protein being α-amylase. The mRNA for α-amylase has been further purified by dimethylsulfoxide-formamide-sucrose density gradient centrifugation and by gel electrophoresis. By these methods, its molecular weight has been determined to be 580,000.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bailey J. M., Davidson N. Methylmercury as a reversible denaturing agent for agarose gel electrophoresis. Anal Biochem. 1976 Jan;70(1):75–85. doi: 10.1016/s0003-2697(76)80049-8. [DOI] [PubMed] [Google Scholar]
  2. Blobel G., Dobberstein B. Transfer of proteins across membranes. I. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma. J Cell Biol. 1975 Dec;67(3):835–851. doi: 10.1083/jcb.67.3.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  4. Bruening G., Beachy R. N., Scalla R., Zaitlin M. In vitro and in vivo translation of the ribonucleic acids of a cowpea strain of tobacco mosaic virus. Virology. 1976 Jun;71(2):498–517. doi: 10.1016/0042-6822(76)90377-9. [DOI] [PubMed] [Google Scholar]
  5. Chrispeels M. J., Varner J. E. Gibberellic Acid-enhanced synthesis and release of alpha-amylase and ribonuclease by isolated barley and aleurone layers. Plant Physiol. 1967 Mar;42(3):398–406. doi: 10.1104/pp.42.3.398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ho D. T. Response of barley aleurone layers to abscisic Acid. Plant Physiol. 1976 Feb;57(2):175–178. doi: 10.1104/pp.57.2.175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ho D. T., Varner J. E. Hormonal control of messenger ribonucleic acid metabolism in barley aleurone layers. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4783–4786. doi: 10.1073/pnas.71.12.4783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jacobsen J. V. Interactions between Gibberellic Acid, Ethylene, and Abscisic Acid in Control of Amylase Synthesis in Barley Aleurone Layers. Plant Physiol. 1973 Jan;51(1):198–202. doi: 10.1104/pp.51.1.198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jacobsen J. V., Zwar J. A. Gibberellic acid causes increased synthesis of RNA which contains poly(A) in barley aleurone tissue. Proc Natl Acad Sci U S A. 1974 Aug;71(8):3290–3293. doi: 10.1073/pnas.71.8.3290. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Okita T. W., Decaleya R., Rappaport L. Synthesis of a possible precursor of alpha-amylase in wheat aleurone cells. Plant Physiol. 1979 Jan;63(1):195–200. doi: 10.1104/pp.63.1.195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  13. Taiz L., Honigman W. A. Production of cell wall hydrolyzing enzymes by barley aleurone layers in response to gibberellic Acid. Plant Physiol. 1976 Sep;58(3):380–386. doi: 10.1104/pp.58.3.380. [DOI] [PMC free article] [PubMed] [Google Scholar]

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