Monitoring intracellular protein profiles with two-dimensional gel electrophoresis.
Two-dimensional polyacrylamide gel electrophoresis (2D PAGE) is a method of separating complex protein mixtures, such as whole cell extracts, on the basis of protein isoelectric point and molecular weight. In bioprocess engineering, conventional 2D PAGE has tremendous potential to yield detailed information on the intracellular effect of various process conditions. It has been used in our work to examine global intracellular changes occurring in a typical cycloheximide fermentation and to look at the feedback regulatory behavior of cycloheximide biosynthesis. Application of the technique for bioprocess monitoring will require that the time necessary for preparation of a 2D electropherogram be substantially shortened. This may be accomplished by performing the separation on a miniature scale or eventually by use of capillary electrophoresis for one or more of the separations. Advantages and disadvantages of these two approaches are discussed.
Biomedical Engineering and Instrumentation Program, National Institutes of Health, Bethesda, Maryland 20892.
K H
KH
Dykstra
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HY
Wang
D003513:Cycloheximide / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D007526:Isoelectric Point; D008970:Molecular Weight; D011506:Proteins / Q000302:isolation & purification*; D013305:Streptomyces griseus / Q000378:metabolism
D011506:Proteins; D003513:Cycloheximide