ABSTRACT
There was a linear increase in poly (A+) polymerase activity in the C. arietinum epicotyls during germination. Six-day-old auxin treated seedlings showed about 3-4 fold stimulation in enzyme activity, accompanied with 3- fold rise in the relative abundance of poly (A+) RNA levels. Actinomycin D, cycloheximide, cordycepin and amino acid analogues caused dramatic decline in poly (A+) polymerase as well as poly (A+) RNA levels. It seems that auxin induced a de novo synthesis of this enzyme.
Subject(s)
Amino Acids/metabolism , Antineoplastic Agents/pharmacology , Cicer/drug effects , Cycloheximide/pharmacology , Dactinomycin/pharmacology , Deoxyadenosines/pharmacology , Electrophoresis, Polyacrylamide Gel , Gene Expression Regulation, Plant , Germination , Indoleacetic Acids/pharmacology , Poly A/metabolism , Polynucleotide Adenylyltransferase/isolation & purification , Protein Synthesis Inhibitors/pharmacology , RNA/metabolism , Seeds/drug effectsABSTRACT
A simple and inexpensive shaker/Erlenmeyer flask system for largescale cultivation of insect cells is described and compared to a commercial spinner system. On the basis of maximum cell density, average population doubling time and overproduction of recombinant protein, a better result was obtained with a simpler and less expensive biorector consisting of Erlenmeyer flasks and an ordinary shaker waterbath. Routinely, about 90 mg of pure poly(ADP-ribose) polymerase catalytic domain was obtained for a total of 3 x 10(9) infected cells in three liters of culture.