Study On Cloning, Expression, Purification Of Mouse Dentin Sialophosphoprotein; Preparation And Specificity Study Of Anti-Mouse Dentin Sialophosphoprotein Antibodies

The molecular basis underlying the mineralization process associated with the conversion from predentin to dentin is poorly known. What is clear that two dentin specific protein, dentin phosphoprotein (DPP), the most abundant Non-collagenous protein (NCP) in dentin, and dentin sialoprotein (DSP), but only accounting for 5-8% of the weight of dentin NCPs, secreted by odontoblasts and appearing at the mineralization front, are believed to participate in the nucleation process and in hydroxyapatite growth during dentin formation. Recent findings showed that both DPP and DSP cDNA were present in the same transcript and the same DSPP arrangement was found at the genomic level. Furthermore, DSP-DPP was found to exist as a continuous open reading frame in both mouse and rat. These findings prompted us to investigate whether the mature DSP and DPP protein in dentin are cleavage products derived from the continuous DSPP open reading frame and whether a single DSPP protein exists. To answer these questions, we used the molecular biological techniques to clone and partially sequence the mouse DPP and DSPP, and express in E. coli and purify by Ni-NTA resin. Then an anti-DSPP antibody is prepared to proceed the Western blot and immunohistochemical study. This paper includes following 3 parts:1. Cloning And Partially Sequencing Of Mouse DPP and DSPP...