OPTIMIZATION OF POLYETHYLENE-GLYCOL MEDIATED TRANSIENT GENE-EXPRESSION IN PEA PROTOPLASTS

Authors
NICOLAISEN M POULSEN GB
Citation
M. Nicolaisen et Gb. Poulsen, OPTIMIZATION OF POLYETHYLENE-GLYCOL MEDIATED TRANSIENT GENE-EXPRESSION IN PEA PROTOPLASTS, Plant cell, tissue and organ culture, 35(1), 1993, pp. 93-97
Citations number
25
Categorie Soggetti
Plant Sciences
Journal title
Plant cell, tissue and organ culture → ACNP
ISSN journal
01676857
Volume
35
Issue
1
Year of publication
1993
Pages
93 - 97
Database
ISI
SICI code
0167-6857(1993)35:1<93:OOPMTG>2.0.ZU;2-V
Abstract
We have investigated factors influencing polyethylene glycol mediated DNA uptake and beta-glucuronidase expression in pea (Pisum sativum L.) protoplasts. It was found that for optimal beta-glucuronidase express ion the molecular weight and concentration of polyethylene glycol shou ld be 4000 and 20%, respectively. The amount of plasmid DNA should be 25 mug per 5 x 10(5) protoplasts in each treatment, and the concentrat ion of Mg2+ in the transformation buffer should be 15 mM. The optimize d protocol was applicable to all four pea cultivars tested.