title: Aggregation of lysine-containing zeins into protein bodies in Xenopus oocytes.
abstract: Zeins, the storage proteins of maize, are totally lacking in the essential amino acids lysine and tryptophan. Lysine codons and lysine- and tryptophan-encoding oligonucleotides were introduced at several positions into a 19-kilodalton zein complementary DNA by oligonucleotide-mediated mutagenesis. A 450-base pair open reading frame from a simian virus 40 (SV40) coat protein was also engineered into the zein coding region. Messenger RNAs for the modified zeins were synthesized in vitro with an SP6 RNA polymerase system and injected into Xenopus laevis oocytes. The modifications did not affect the translation, signal peptide cleavage, or stability of the zeins. The ability of the modified zeins to assemble into structures similar to maize protein bodies was assayed by two criteria: assembly into membrane-bound vesicles resistant to exogenously added protease, and ability to self-aggregate into dense structures. All of the modified zeins were membrane-bound; only the one containing a 17-kilodalton SV40 protein fragment was unable to aggregate. These findings suggest that it may be possible to create high-lysine corn by genetic engineering.
authors.affiliation: Department of Botany and Plant Pathology, Purdue University, West Lafayette, IN 47907.
authors.forename: J C
authors.initials: JC
authors.lastname: Wallace
authors.forename: G
authors.initials: G
authors.lastname: Galili
authors.forename: E E
authors.initials: EE
authors.lastname: Kawata
authors.forename: R E
authors.initials: RE
authors.lastname: Cuellar
authors.forename: M A
authors.initials: MA
authors.lastname: Shotwell
authors.forename: B A
authors.initials: BA
authors.lastname: Larkins
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D004247:DNA / Q000235:genetics; D004274:DNA, Recombinant; D005260:Female; D005818:Genetic Engineering; D008239:Lysine / Q000235:genetics; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009154:Mutation; D009865:Oocytes / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D013539:Simian virus 40 / Q000235:genetics; D014982:Xenopus laevis; D003313:Zea mays; D015028:Zein / Q000235:genetics / Q000378:metabolism*
chemical_list: D004274:DNA, Recombinant; D046911:Macromolecular Substances; D012333:RNA, Messenger; D004247:DNA; D015028:Zein; D010447:Peptide Hydrolases; D008239:Lysine