Aszódi A, Beier D R, Hiripi L, Bösze Z, Fässler R
Department of Experimental Pathology, University Hospital, Lund, Sweden.
Matrix Biol. 1998 Mar;16(9):563-73. doi: 10.1016/s0945-053x(98)90067-1.
The mouse cartilage matrix protein gene (Crtm) was isolated from a cosmid library using a mouse Crtm cDNA fragment as probe. Crtm spans 12.2 kb from the start of translation to the polyadenylation signal sequence and comprises eight exons. Sequencing of the 1.9 kb 5' flanking region revealed a TATA-like box 72 bp upstream from the initiator Met codon as well as several cis-acting motifs known to bind eukaryotic transcription factors. Analysis of the exon-intron junctions demonstrated that the last intron does not follow the gt/ag rule but belongs to the minor class of pre-mRNA introns that contain "at" and "ac" at their 5'and 3' ends, respectively. Single-strand conformation polymorphism analysis was used to map Crtm to the distal part of chromosome 4 between the microsatellite markers D4Mit16 and D4Mit339. Achodroplasia (cn), a recessive skeletal disorder in mice, has already been mapped to this region. Immunostaining for CMP and sequence of Crtm in cn/cn mice failed to reveal any disease-specific mutations, suggesting that mutations in Crtm do not cause achondroplasia.
利用小鼠软骨基质蛋白基因(Crtm)的cDNA片段作为探针,从黏粒文库中分离出小鼠软骨基质蛋白基因(Crtm)。Crtm从翻译起始点到聚腺苷酸化信号序列跨度为12.2 kb,由八个外显子组成。对1.9 kb的5'侧翼区域进行测序,发现在起始甲硫氨酸密码子上游72 bp处有一个类似TATA盒的结构,以及几个已知能结合真核转录因子的顺式作用基序。对外显子-内含子连接区的分析表明,最后一个内含子不遵循gt/ag规则,而是属于前体mRNA内含子的小类,其5'和3'末端分别含有“at”和“ac”。利用单链构象多态性分析将Crtm定位到4号染色体远端微卫星标记D4Mit16和D4Mit339之间的区域。小鼠的一种隐性骨骼疾病——软骨发育不全(cn)已被定位到该区域。对cn/cn小鼠进行CMP免疫染色和Crtm测序,未发现任何疾病特异性突变,这表明Crtm中的突变不会导致软骨发育不全。