An analysis of the relative activities of a number of promoter constructs from genes which are expressed during late pollen development as determined by particle bombardment
1995
Lonsdale, D.M. | Allen, R.L. | Belostotsky, D. | Ghose, T.K. | Harvey, A.J. | Rogers, H.J. | Tebbut, S.J. | Trick, M.
The promoters of a tobacco actin gene, a tobacco pectate lyase, a tobacco and maize polygalacturonase and a Brassica S-locus related gene have been fused to the beta-glucuronidase reporter gene and their activities determined by biolistic transient assay in tobacco pollen. In stably transformed tobacco all the transgenes with the exception of Cauliflower Mosaic Virus-35S-beta-glucuronidase appear to express efficiently in maturing pollen. Transient assay analysis showed that the tobacco pectate lyase and the polygalacturonase constructs were 8X more active than the tobacco actin construct, and that the tobacco polygalacturonase construct was some 33X more active than the maize polygalacturonase construct. Constructional manipulations that altered the lengths of the 5'-untranslated leaders including one which resulted in the removal of a 490 bp leader intron had little effect on the observed level of expression. However, the alteration of the context of the ATG from A/TnnATGG to CnnATGT resulting in a 70% reduction in the observed levels of activity, was obtained with the pectate lyase and polygalacturonase promoters. An identical reductional was also observed in transgenic plant populations transformed with the polygalacturonase transgenes.
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