Canadian Forest Service Publications

Microprojectile-DNA delivery in conifer species: factors affecting assessment of transient gene expression using the ß-glucuronidase reporter gene. 1993. Charest, P.J.; Caléro, N.; Lachance, D.A.; Datla, R.S.S.; Duchesne, L.C.; Tsang, E.W.T. Plant Cell Reports 12: 189-193.

Year: 1993

Issued by: National Capital Region

Catalog ID: 10740

Language: English

Availability: PDF (request by e-mail)

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The Biolistic® microprojectile DNA-delivery method was used to test the usefulness in conifers of eight gene consructs based on the 35S promoter, the AMV translational enhancer, and gene fusion between the -glucuronidase and the neomycin phosphotransferase II genes. The evaluation was done with embryogenic cells of Picea glauca, where the relative strengths of the promoters were 35S-35S-AMVE>35S-AMVE>35S-35S>35S as evaluated by transient gene expression. The fusion gene of GUS and NPT II gave lower levels of transient gene expression than the unfused GUS gene as detected by X-GLU histochemical assays. Experiments comparing the EM promoter of wheat and the 35S-35S-AMVE promoter (with and without fusion between GUS and NPT II) were done in Picea rubens, P. mariana, P. glauca, and Larix x eurolepis. The unfused gene with the 35S-35S-AMVE promoter gave higher levels of transient gene expression than the fused GUS-NPT II gene. The fluorescent MUG assay was more sensitive than the histochemical X-GLU assay to detect the activity of the -glucuronidase gene.