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DNA binding sites recognised in vitro by a knotted class 1 homeodomain protein encoded by the hooded gene, k , in barley ( Hordeum vulgare )
Author(s) -
Krusell Lene,
Rasmussen Inge,
Gausing Kirsten
Publication year - 1997
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(97)00382-7
Subject(s) - fusion protein , gene , complementary dna , dna , homeobox , biology , maltose binding protein , fusion gene , hordeum vulgare , genetics , microbiology and biotechnology , dna binding protein , transcription factor , biochemistry , recombinant dna , botany , poaceae
The homeodomain of the knotted classes of transcription factors from plants differs from the well characterized Antp/En type homeodomains from Drosophila at key amino acid residues contributing to the DNA binding. A cDNA, Hvh21 , derived from the hooded gene and encoding a full length homolog of knotted1 from maize was isolated from barley seedlings and expressed as a maltose binding protein fusion in E. coli . The purified HvH21‐fusion protein selected DNA fragments with 1–3 copies of the sequence TGAC. Gel shift experiments showed that the TGAC element was required for binding and the results further indicate that the HvH21‐fusion protein binds DNA as a monomer.