Transient gene expression in cassava using high-velocity microprojectiles.
The bacterial gene encoding beta-glucuronidase (GUS) was transiently expressed in cassava leaves following the introduction of the gene by microparticle bombardment. The DNA expression vector used to introduce the reporter gene is a pUC 19 derivative and consisted of a CaMV 35S promoter (P35S), the GUS coding region and 7S polyadenylation region. Several other promoters and regulating sequences were tested for efficiency in cassava leaves. Two derivatives of the P35S, one including a partial duplication of the upstream region of the P35S and the other containing a tetramer of the octopine synthase enhancer, were found to be expressed at three times the level of the P35S in cassava leaves. The ubiquitin 1 promoter from Arabidopsis thaliana was expressed at the same level as the P35S. No influence on the level of expression was observed when different 3' ends were used. The biolistic transient gene expression system in cassava leaves allows rapid analysis of gene constructs and can serve as a preliminary screen for chimeric gene function in the construction of transgenic cassava plants.
Franche
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ORSTOM, Saint Louis, MO.
Bogusz
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Schöpke
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Fauquet
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Beachy
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RN
D003001:Cloning, Molecular / Q000379:methods*; D015870:Gene Expression; D005966:Glucuronidase / Q000235:genetics; D002365:Manihot / Q000235:genetics*; D009029:Mosaic Viruses / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014414:Tungsten; D014452:Ubiquitins / Q000235:genetics
D014452:Ubiquitins; D005966:Glucuronidase; D014414:Tungsten