Expression of the sorghum 10-member kafirin gene cluster in maize endosperm.

Publisher: Oxford University Press Country of Publication: England NLM ID: 0411011 Publication Model: Electronic Cited Medium: Internet ISSN: 1362-4962 (Electronic) Linking ISSN: 03051048 NLM ISO Abbreviation: Nucleic Acids Res Subsets: MEDLINE

Publication: 1992- : Oxford : Oxford University Press
Original Publication: London, Information Retrieval ltd.

Transformation, Genetic*; Plant Proteins/*genetics ; Plants, Genetically Modified/*genetics ; Zea mays/*genetics; Chromatin/chemistry ; Chromosomes, Artificial, Bacterial ; Gene Expression ; Gene Expression Regulation, Plant ; Genome, Plant ; Multigene Family ; Plant Proteins/metabolism ; Plants, Genetically Modified/metabolism ; Prolamins ; Sorghum/genetics ; Zea mays/metabolism

Functional analysis of chromosomal segments containing linked genes requires the insertion of contiguous genomic sequences from bacterial artificial chromosomes (BACs) into the genome. Therefore, we introduced a 90-kb large BAC clone carrying a 10-copy tandem array of kafirin storage protein genes from sorghum linkage group J, mixed with a selectable marker gene, directly into maize cells using the particle bombardment method. Transgenic plants were regenerated and seeds from eight different transgenic lines were produced. One such transgenic plant was selected that had the entire kafirin gene cluster on a single continuous DNA fragment spanning more than 45 kb integrated into its genome. When alcohol-soluble proteins from individual T2 and T3 seeds of this event were analyzed, significant levels of kafirin were found in addition to the endogenous zein storage proteins, demonstrating that the large exogenous DNA segment is stably integrated into the maize genome and expressed at high levels in subsequent generations. Therefore, we could provide a new utility of plant transformation by the particle bombardment method for functional genomics of multigene families and the modification of the nutritive quality of cereal grains. Despite a tandem array of highly homologous sequences at the transgenic locus, no gene silencing was observed, probably owing to the effects of co-transformed flanking sequences. The expression studies of the transgenic locus also revealed new features of storage protein gene promoters that differed from previous transient gene expression studies, thereby illustrating the significance of the concentration and configuration of DNA-protein interactions in the regulation of gene expression.

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0 (Chromatin)
0 (Plant Proteins)
0 (Prolamins)
0 (kafirin protein, Sorghum bicolor)

Date Created: 20041231 Date Completed: 20050119 Latest Revision: 20181113

20231215

PMC545481

10.1093/nar/gnh183

15625231