RESUMEN
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.
Asunto(s)
Aminoácidos/metabolismo , Antineoplásicos/farmacología , Cicer/efectos de los fármacos , Cicloheximida/farmacología , Dactinomicina/farmacología , Desoxiadenosinas/farmacología , Electroforesis en Gel de Poliacrilamida , Regulación de la Expresión Génica de las Plantas , Germinación , Ácidos Indolacéticos/farmacología , Poli A/metabolismo , Polinucleotido Adenililtransferasa/aislamiento & purificación , Inhibidores de la Síntesis de la Proteína/farmacología , ARN/metabolismo , Semillas/efectos de los fármacosRESUMEN
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.