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Title: Evaluation of promoters and visual markers for transformation of eastern white pine

Author: Zipf, Allan; Diner, Alex M.; Ward, Rufina; Bharara, Suman; Brown, George Jr.; Nagmani, R.; Pareek, L.K.; Sharma, Govind C.;

Date: 2001

Source: New Forests 21: 45-58, 2001.

Publication Series: Miscellaneous Publication

Description: This report serves to evaluate possible promoters for use in the production of trmsgenic eastern white pine (Pinus strobus L.). Embryogenic cultures of eastern white pine were bombarded with gold particles coated separately with a variety of gene constructs containing the Uida B-glucoronidase (GUS) or green flourescent protein (GFP) reporter gene. Transient expression of the UidA gene. driven by a novel algal virus adenine methyl transferuse gene promoter. as well as live other promoters used in angiosperm transformation. were evaluated. The maize alcohol-dehydrogenase promoter was not effective in eastern white pine cultures. The constrwct with the doubled Cauliflower Mosaic Virus 35S promoter plus Alfalfa Mosaic Virus enhancer showed the highest levels of expression. GUS expression was detected within 24 hours, but decreased after 5 days and was not detectable 15 days after bombardment. Expression of GUS activity was recorded mainiy in sorrutic embryonal heads of various stages of development and occasionally in suspensor cells. Similar to GUS expression. moditied green Huoresccnt protein (GFP) was detected in the embryonul head ceils 7-4 hours after bombardment. GFP-expressing suspensor cells were both more infrequent and difficult to detect, as their highly vacuolate nature rendered the GFP presence less visible against the yellow background autoflourescence.

Keywords: biolistics, GFP-GUS, Pinus strubus L., transient gene expression

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Zipf, Allan; Diner, Alex M.; Ward, Rufina; Bharara, Suman; Brown, George, Jr.; Nagmani, R.; Pareek, L.K.; Sharma, Govind C. 2001. Evaluation of promoters and visual markers for transformation of eastern white pine. New Forests 21: 45-58, 2001.


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