RESUMO
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.
Assuntos
Aminoácidos/metabolismo , Antineoplásicos/farmacologia , Cicer/efeitos dos fármacos , Cicloeximida/farmacologia , Dactinomicina/farmacologia , Desoxiadenosinas/farmacologia , Eletroforese em Gel de Poliacrilamida , Regulação da Expressão Gênica de Plantas , Germinação , Ácidos Indolacéticos/farmacologia , Poli A/metabolismo , Polinucleotídeo Adenililtransferase/isolamento & purificação , Inibidores da Síntese de Proteínas/farmacologia , RNA/metabolismo , Sementes/efeitos dos fármacosRESUMO
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.