Transposon tagging of the maize Glossy2 locus with the transposable element En/Spm.
The Glossy2 (Gl2) locus of maize is required for the formation of the epicuticular wax layer of young plants. gl2 mutant seedlings can be visually identified because of their glossy leaf surface which is different from the dull surface of wild-type seedlings. The Gl2 locus was isolated by transposon tagging. Seven unstable mutations, gl2-m2 to gl2-m8, were induced in a parental strain carrying an active transposable Activator (Ac) element in the unstable wx-m7 allele. Genetic tests on the gl2-m2 allele indicated that it was not caused by the Ac element but by the insertion of the transposable element Enhancer/Suppressor-Mutator (En/Spm). A Sa/l restriction fragment segregating with the mutant phenotype was identified, by Southern analysis, using sequences from the En/Spm element as a probe. Part of the fragment was cloned and was shown to carry part of the unstable gl2-m2 allele. These gl2 sequences were used to identify a genomic fragment carrying the wild-type allele and to isolate its corresponding cDNA sequence. The predicted Glossy2 protein consists of 426 amino acids. No similar amino acid sequence was found in protein data banks and the biochemical function of the Gl2 gene product is still unknown. The wild-type Gl2 transcript is found predominantly in juvenile leaves. The transcript level in the leaves of seedlings homozygous for a stable recessive gl2-ref allele is hardly detectable.
Tacke
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E
Istituto Sperimentale per la Cerealicoltura, Sezione di Bergamo, Italy.
Korfhage
C
C
Michel
D
D
Maddaloni
M
M
Motto
M
M
Lanzini
S
S
Salamini
F
F
Döring
H P
HP
D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D017931:DNA Primers; D004251:DNA Transposable Elements; D018076:DNA, Complementary; D018744:DNA, Plant / Q000302:isolation & purification / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific; D004742:Enhancer Elements, Genetic; D008969:Molecular Sequence Data; D010641:Phenotype; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics*; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping; D013489:Suppression, Genetic; D014158:Transcription, Genetic; D003313:Zea mays / Q000235:genetics* / Q000378:metabolism
D017931:DNA Primers; D004251:DNA Transposable Elements; D018076:DNA, Complementary; D018744:DNA, Plant; C098135:Gl2 protein, Zea mays; D010940:Plant Proteins; D015252:Deoxyribonucleases, Type II Site-Specific; C029792:GTCGAC-specific type II deoxyribonucleases