Isolation of markers for chondro-osteogenic differentiation using cDNA library subtraction - Molecular cloning and characterization of a gene belonging to a novel multigene family of integral membrane proteins
Isolation of markers for chondro-osteogenic differentiation using cDNA library subtraction - Molecular cloning and characterization of a gene belonging to a novel multigene family of integral membrane proteins
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DOI：
10.1074/jbc.271.32.19475
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发表时间：
1996-08-09
影响因子：
4.8
通讯作者：
Merregaert, J
中科院分区：
文献类型：
--
作者：
Deleersnijder, W;Hong, GZ;Merregaert, J
为了寻找与软骨成骨分化相关的新的标记分子，我们建立了一个基于cDNA文库消减的差异筛选系统，从体外培养的髁突构建的cDNA文库中分离到一个新的基因，命名为E25，E25全长cDNA编码263个氨基酸残基的II型膜整合蛋白。原位杂交实验表明，E25表达于生后髁突软骨外膜边缘，其它软骨组织及其软骨膜均为阴性。在牙胚和毛囊中成熟骨的骨小梁上也检测到较强的原位杂交信号，北方印迹分析显示，E25在成骨组织，如新生小鼠颅骨、爪、尾和皮肤中强表达。该表达谱表明，E25可能是软骨成骨分化的有用标志物。DNA同源性分析表明，E25属于一个新的多基因家族，在人和小鼠中都有三个成员。将小鼠E25基因位点（Itm2）定位于X染色体。
To identify novel marker molecules associated with chondro-osteogenic differentiation, we have set up a differential screening system based on a cDNA library subtraction in organ cultures of prenatal mouse mandibular condyles, Differential screening of a cDNA library constructed from in vitro cultured condyles allowed the isolation of a novel gene, named E25, Full-length E25 cDNA is predicted to encode a type II integral membrane protein of 263 amino acid residues, In situ hybridization experiments show that E25 is expressed in the outer perichondrial rim of the postnatal mandibular condyle, which contains the proliferating progenitor cells, but not in the deeper layers of the condyle containing the more differentiated chondroblasts and chondrocytes, Other cartilagenous tissues and their perichondrium were negative, Strong in situ hybridization signals were also detected on bone trabeculae of mature bone in tooth germs and in hair follicles, Northern blot analysis showed strong expression in osteogenic tissues, such as neonatal mouse calvaria, paws, tail, and in skin, This expression profile suggests that E25 could be a useful marker for chondro-osteogenic differentiation. Homology searches of DNA databanks showed that E25 belongs to a novel multigene family, containing three members both in man and mouse. The mouse E25 gene locus (Itm2) was mapped to the X chromosome.