Anti-MBD1 monoclonal antibody, clone 3I4

Mouse beta defensin-1 (mBD-1) is a cationic peptide with broad antimicrobial activity. The mBD-1 gene was cloned and fused with TrxA to construct pET32-mBD1, which was transformed into E. coli BL21 (DE3). The optimal expression conditions of fusion protein TrxA-mBD1 were: cultivation at 32A degrees C in 2 x YT medium, induction with 0.2 mM isopropylthio--galactoside (IPTG), and post-induction expression for 8 h. The fusion protein was highly soluble (90.0%) and accounted for 65% of the total soluble protein; and its volumetric productivity reached 0.67 g/l, i.e., 0.14 g/l of recombinant mBD-1. At 5 mu M, purified recombinant mBD-1 killed 50% of Candida albicans.

MeCP2 is a transcription factor that recognizes and binds symmetrically methylated CpG dinucleotides to repress transcription. MeCP2 can associate with the Sin3a/histone deacetylase corepressor complex and mediate repression in a histone deacetylase-dependent manner. In extracts from rodent tissues, cultured cells, and Xenopus laevis oocytes, we find that only a small amount of mammalian MeCP2 interacts with Sin3a and that this interaction is not stable. Purification of rat brain MeCP2 (53 kDa) indicates no associated proteins despite an apparent molecular mass by size exclusion chromatography of 400-500 kDa. Biophysical analysis demonstrated that the large apparent size was not because of homo-multimerization, as MeCP2 consistently behaves as a monomeric protein that has an elongated shape. Our findings indicate the MeCP2 is not an obligate component of the Sin3a corepressor complex and may therefore engage a more diverse range of cofactors for repressive function.