| First Author | de Jong R | Year | 1992 |
| Journal | Gene | Volume | 116 |
| Issue | 2 | Pages | 195-203 |
| PubMed ID | 1353046 | Mgi Jnum | J:2103 |
| Mgi Id | MGI:50627 | Doi | 10.1016/0378-1119(92)90516-r |
| Citation | de Jong R, et al. (1992) DNA-binding activity of the murine homeodomain protein Hox-2.3 produced by a hybrid phage T7/vaccinia virus system. Gene 116(2):195-203 |
| abstractText | Homeobox-containing genes encode transcription factors that, via the homeodomain, bind specifically to DNA. To study the DNA-binding properties of the murine homeodomain-containing protein, Hox-2.3, a hybrid expression system was used, combining gene expression by recombinant vaccinia virus (reVV) with bacteriophage T7 transcription. Expression was achieved by co-infecting HeLa cells with two reVVs, one expressing the T7-RNA polymerase-encoding gene directed by the VV promoter, P7.5, and another containing the Hox-2.3 coding sequence under control of a T7 promoter [Fuerst et al., Mol. Cell. Biol. 7 (1987) 2538-2544]. Co-infected HeLa cells produced large amounts of full-length Hox-2.3 protein. Cytoplasmic and nuclear extracts from these cells were used to examine DNA-binding specificity in vitro. reVV-produced Hox-2.3 protein bound to oligos that contained one or several copies of the common homeodomain-binding site, 5'-TCA-ATTAAAT, and to a lesser extent to multiple (TAA) repeats. Using Southwestern blot analysis, no Hox-2.3-binding sites were detected in a region of the Hox-2 cluster containing the Hox-2.3, Hox-2.4 and Hox-2.5 genes. |
