Abstract
Primer extension was used to characterize α-amylase mRNAs from aleurone tissue of barley (Hordeum vulgare L. cv. Himalaya) grains. Two synthetic oligonucleotides, specific for the low-pI and high-pI α-amylase groups, were used as primers for synthesis of cDNA from total RNA preparations. Between them, these two oligonucleotides appear to account for all major α-amylase mRNAs as judged by hybrid-arrested translation of α-amylase mRNAs in a cell-free system.
Reconstruction experiments indicated that the levels of extended primers (determined by scintillation counting) were directly proportional to the level of input mRNA over a wide range. This indicates that the technique is suitable for quantification of relative levels of individual α-amylase from approximately 2% to 100% of maximal levels.
The nucleotide sequences of extended primers defined two different α-amylase mRNAs in each of the low-pI and high-pI groups, and possibly a third mRNA in the high-pI group.
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Chandler, P.M., Huiet, L. Primer extension studies on α-amylase mRNAs in barley aleurone. I. Characterization and quantification of the transcripts. Plant Mol Biol 16, 627–635 (1991). https://doi.org/10.1007/BF00023427
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DOI: https://doi.org/10.1007/BF00023427