Expression pattern of Dentin Matrix Protein 1 (DMP1) during Development via EGFP Reporter Gene and Construction of DMP1 promoter-Shh cDNA Over-expression Vector
Date Issued
2007
Date
2007
Author(s)
Lin, Li-Fen
DOI
zh-TW
Abstract
In the process of osteogenesis and dentinogenesis, osteoblasts and odontoblasts secrete non-collagenous proteins to form extracellular matrix (ECM). One group of the non-collagenous proteins is SIBLING family (SIBLING family; Small-Integrin-Binding Ligand, N-linked Glyco- protein). DMP-1 (Dentin Matrix Protein 1; DMP-1) is a member of the family.
DMP-1, an acidic phosphorylated protein, is expressed biofunctionally in bone and teeth calcification. Previous studies have shown that DMP-1 is expressed in odontoblasts, ameloblsats, cementoblasts, calvaeia, and long bone during teeth and bone differentiation.
Upstream 2,783 Kb fragment of human DMP-1 promoter is functional by the expression of the reporter gene (EGFP, enhanced green fluorescent protein) in transgenic zabrafish and mice (Yu, 2004). In this study, I found that Hu-DMP-1-GFP-2.783K-36 can drive the expression of the reporter gene (EGFP) in lymph node, palm, hair follicle, maxilla, mandible, and tail at the stage of 15.5 dpc (day post coitum). Moreover, Hu-DMP- 1-GFP-2.783K-36 also can drive the expression of the reporter gene (EGFP) in the bone, trachea, thymus, spleen, kidney, peyer’s patch, testis, stomach, esophagus and tongue in the transgenic mice at the postnatal development (one month). The overall results suggest that location of the DMP-1 is not restricted to mineralized tissues and may be present in non-mineralized tissue. Furthermore, the expression pattern of DMP1 was further confirmed by RT-PCR and in situ hybridization.
I have constructed the pDMP-shh-IRES-hrGFP chimeric gene, and the constructs were micro-injected into the zebrafish eggs. In this way, Shh can be over-expressed in the bone, and simultaneously the transgenic fish containing the construct can be screened by the expression of GFP. However, no transgenic stable line was obtained.
Subjects
牙本質基質蛋白
反轉錄聚合酶
鏈鎖反應
原位雜合法
牙齒
骨頭
綠色螢光蛋白
DMP-1
RT-PCR
In situ hybridization
teeth
bone
GFP
Type
other