Production of mouse Hox-2.1 protein in Escherichia coli: characterisation of in vitro binding to DNA.
The developmentally regulated mouse Hox-2.1 gene encodes a homeodomain-containing (Hox) protein which is likely to function as a transcription factor. We expressed the DNA coding for full-length Hox-2.1 protein in a T7 promoter-containing vector in bacteria, which produced low levels of protein showing weak DNA-binding activity. Synthesis of a truncated polypeptide lacking all the sequence upstream from the homeodomain enabled us to produce greater amounts of protein and demonstrate its sequence-dependent DNA binding. The tetranucleotide ATTA is necessary for binding, but a single copy is not by itself sufficient. Flanking sequences are important; in particular a cytosine immediately 5' to the ATTA enhances binding.
MRC Human Genetics Unit, Western General Hospital, Edinburgh, Scotland.
P S
PS
Budd
I J
IJ
Jackson
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D003854:Deoxyribonucleotides; D004926:Escherichia coli / Q000235:genetics*; D018398:Homeodomain Proteins; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids; D014176:Protein Biosynthesis; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D003854:Deoxyribonucleotides; D018398:Homeodomain Proteins; C053925:Hoxb5 protein, mouse