Premium
A rapid and simple method of producing high‐resolution zymograms from Brassica and related species on isoelectric‐focussing gels
Author(s) -
O'Neill C. M.,
Mathiasu R. J.
Publication year - 1995
Publication title -
plant breeding
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.583
H-Index - 71
eISSN - 1439-0523
pISSN - 0179-9541
DOI - 10.1111/j.1439-0523.1995.tb00787.x
Subject(s) - biology , isoelectric focusing , brassica , isozyme , starch , chromatography , polyacrylamide gel electrophoresis , isoelectric point , gel electrophoresis , electrophoresis , botany , biochemistry , enzyme , chemistry
Isozymes are potentially valuable genetic markers in the identification of hybrid plants, the assessment of seed purity, and in molecular and conventional genetic maps. The starch gels commonly used to prepare Brassica zymograms have very low resolutions. Consequently, isoelectric‐focussing polyacrylamide‐gel electrophoresis was explored as a means of producing high‐resolution zymograms of four Brassica isozymes. Crude sap extracts were found to be unsuitable for electrophoresis. Centrifugation, passage over sepharose beads and overnight dialysis were tested as alternative means of clarifying samples. The zymograms presented here demonstrate that isoelectric focussing, following overnight sample dialysis, results in zymograms which, in comparison to their starch counterparts, have vastly improved separation and resolution.