Details

Autor(en) / Beteiligte

• Chandler, P.M
• Huiet, L

Titel

Primer extension studies on alpha-amylase mRNAs in barley aleurone. I. Characterization and quantification of the transcripts

Ist Teil von

• Plant molecular biology, 1991-04, Vol.16 (4), p.627-635

Ort / Verlag

Dordrecht: Springer

Erscheinungsjahr

1991

Quelle

SpringerLINK Contemporary (Konsortium Baden-Württemberg)

Beschreibungen/Notizen

• Primer extension was used to characterize alpha-amylase mRNAs from aleurone tissue of barley (Hordeum vulgare L. cv. Himalaya) grains. Two synthetic oligonucleotides,specific for the low-pl and high-pl alpha-amylase groups, were used as primers for synthesis of cDNAfrom total RNA preparations. Between them, these two oligonucleotides appear to account for all major alpha-amylase mRNAs as judged by hybrid-arrested translation of alpha-amylase mRNAs in a cell-freesystem. Reconstruction experiments indicated that the levels of extendedprimers (determined by scintillation counting) were directly proportional to the level of input mRNAover a wide range. This indicates that the technique is suitable for quantification of relative levelsof individual alpha-amylase from approximately 2% to 100% of maximal levels. The nucleotide sequences of extended primers defined twodifferent alpha-amylase mRNAs in each of the low-pl and high-pl groups, and possibly a third mRNA in thehigh-pl group.