Construction Of The Mouse Enamel Matrix Serine Proteinase Gene Targeting Vector

The mouse enamel matrix serine proteinase(EMSP1)is encoded by PRSS17(serine protease 17)gene. The DNA sequence of EMSP1 is conservative among different species. The mouse and human have six exons and five intros. The mouse PRSS17 gene sequence is 10134bp.Repetitive DNA sequence from the MSR1 family are found in both the mouse and human PRSS17 genes. Like other serine proteinases, EMSP1 use a catalytic triad of serine, histidine and aspartate to hydrolyze bond.There are two classes of proteinases which are responsible for the cleavage of enamel matrix protein. One of them is MMP20 ,and the other is EMSP1.The increasing evidence suggest that EMSP1 plays a key role in the degradation of enamel matrix. The enamel resident proteinases have another role in the enamel development, which may be helpful the proper location of cleavage products. Loss of function resulted from mutation in the genes encoding EMSP1 may inherited amelogenesis imperfect. EMSP1 is so little in enamel matrix of tooth germ, and it is only transcripted and expressed in the transition and early maturation stage. EMSP1 is expressed not only in extremely low amounts in the enamel matrix, but also for a short time during teeth development. This is useful to understand the molecular mechanisms of dental enamel formation. EMSP1 was the first reported proteinase fractionated from the developing enamel matrix. Now the enamel proteinase is the first to be isolated and characterized at both protein and DNA levels.Construction of The mouse enamel matrix serine proteinase Gene Knockout VectorTwo pairs of primers were designed by Oligo 6.0 software and synthesized according to EMSP1 genomic sequence which had sequenced before and PRSS17 sequence (10134) from GenBank. Two gene fragments of...