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Modification of the coding sequence enhances plant expression of insect control protein genes.pdf

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Modification of the coding sequence enhances plant expression of insect control protein genes

Proc. Nati. Acad. Sci. USA Vol. 88, pp. 3324-3328, April 1991 Biochemistry Modification of the coding sequence enhances plant expression of insect control protein genes (Bacilus thuringiensis/insect-resistant plants/synthetic genes) FREDERICK J. PERLAK*, RoY L. FUCHS, DUFF A. DEAN, SYLVIA L. MCPHERSONt, AND DAVID A. FISCHHOFF Plant Sciences, Monsanto Company, 700 Chesterfield Village Parkway, Chesterfield, MO 63198 Communicated by Jeff Schell, January 14, 1991 ABSTRACT Increased expression of the insect control protein genes ofBacilus thunrngiensis in plants has been critical to the development of genetically improved plants with agro- nomically acceptable levels of insect resistance. The expression of the cryIA(b) gene was compared to partially modIfied (3% nucleotide difference) and to fully modified (21% nucleotide difference) crylA(b) and crylA(c) genes in tobacco and tomato. The modified genes increased the frequency of plants that produced the proteins at quantities sufficient to control insects and dramatically increased the levels of these proteins. Among the most highly expressing transformed plants for each gene, the plants with the partially modified crylA(b) gene had a 10-fold higher level ofinsect control protein and plants with the fully modified crylA(b) had a 100-fold higher level of CryIA(b) protein compared with the wild-type gene. Similar results were obtained with the fully modified crylA(c) gene in plants. Specific sequences of the partially modified cryIA(b) gene were analyzed for their ability to affect crylA(b) gene expression in tobacco. The DNA sequence of a single region was idented as important to the improvement of plant expression of the crylA(b) gene. The increased levels of cryIA(b) mRNA were not directly proportional to the increased levels ofCryIA(b) protein in plants transformed with the modified crylA(b) genes, indi- cating that the nucleotide sequence of these genes had an effect in improving their translational efficiency in pla