Transient gene expression in electroporated Picea glauca protoplasts |
| |
Authors: | F. Bekkaoui M. Pilon E. Laine D. S. S. Raju W. L. Crosby D. I. Dunstan |
| |
Affiliation: | (1) Plant Biotechnology Institute, 110 Gymnasium Road, Saskatoon, S7N OW9 Saskatoon, Canada;(2) Present address: Laboratoire de Biologie et Physiologie Végétale, Université de Bordeaux 1, 33405 Talence, France |
| |
Abstract: | The reporter gene for chloramphenicol acetyltransferase (CAT) was introduced into white spruce (Picea glauca (Moench) Voss.) protoplasts by electroporation. CAT transient gene expression was increased by increasing the concentration of pCaMVCN plasmid and was affected by the level of the applied voltage. Highest CAT activities were obtained after electroporation with a pulse of 350V.cm–1 having an exponential decay constant of approximately 105ms. Linearized plasmid constructs gave much higher levels of CAT activity than circular plasmid. Attempts to use the Escherichia coli -glucuronidase gene (-GUS) as a marker gene revealed very high levels of -GUS-like activity in electroporated protoplasts. This activity was mainly due to a small molecule and may mask successful transformation since -GUS-like activity increased when plasmid DNA was present during electroporation.Abbreviations CAT chloramphenicol acetyltransferase - -GUS -glucuronidase - MUG 4-methyl umbelliferyl glucuronide - F microfaradsNRCC No. 29150 |
| |
Keywords: | |
本文献已被 SpringerLink 等数据库收录! |
|