Escribano J, Hernando N, Ghosh S, Crabb J, Coca-Prados M
Department of Ophthalmology and Visual Science, Yale University School of Medicine, New Haven, Connecticut 06510.
J Cell Physiol. 1994 Sep;160(3):511-21. doi: 10.1002/jcp.1041600314.
The non-pigmented ciliary epithelium is largely responsible for the formation of aqueous humor in the mammalian eye. To provide a basis for studies at the molecular level, a directional expression cDNA library was constructed in Uni-ZAP XR vector from poly A+ RNA of the human non-pigmented ciliary epithelial derived ODM-2 cell line. Fifty-three cDNA clones were isolated from the library and characterized by partial sequence analysis. Approximately 49% of the clones exhibited homology with known genes in the GenBank/EMBL databases. The putative identification of these clones may reflect the transcriptional activity of the ODM-2 cells in culture. One of the identified clones, ODM-42-I, was found to be specific and highly expressed in the corneal epithelium. This clone had an exact match with a recently discovered human gene, beta ig-h3 (Skonier et al., 1992, DNA Cell Biol., 11:511-522), which codes a surface recognition protein, inducible by transforming growth factor beta (TGF-beta), and containing a putative binding site (RDG) for integrins. The ODM-42-I cDNA clone displays a distinctive pattern of expression found in the human eye, expressed almost exclusively in the cornea. Further studies, using sera from a synthetic peptide to the carboxy-terminal region of ODM-42-I, reveal that the protein is heterogeneous in charge and is preferentially expressed on the extracellular surface of corneal epithelial cells, and might share immunologic properties with integrins beta 1.
在哺乳动物眼中,无色素睫状上皮在很大程度上负责房水的形成。为了在分子水平上开展研究提供基础,利用人无色素睫状上皮来源的ODM-2细胞系的聚腺苷酸加尾RNA,构建了一个定向表达cDNA文库,该文库克隆于Uni-ZAP XR载体。从该文库中分离出53个cDNA克隆,并通过部分序列分析进行了表征。大约49%的克隆与GenBank/EMBL数据库中的已知基因具有同源性。这些克隆的推定鉴定可能反映了培养中的ODM-2细胞的转录活性。其中一个鉴定出的克隆ODM-42-I,被发现特异性且在角膜上皮中高表达。该克隆与最近发现的一个人类基因βig-h3(Skonier等人,1992年,《DNA与细胞生物学》,11:511 - 522)完全匹配,该基因编码一种表面识别蛋白,可被转化生长因子β(TGF-β)诱导,并且含有一个假定的整合素结合位点(RDG)。ODM-42-I cDNA克隆展示了在人眼中发现的一种独特表达模式,几乎仅在角膜中表达。进一步的研究,使用针对ODM-42-I羧基末端区域的合成肽血清,揭示该蛋白电荷不均一,且优先在角膜上皮细胞的细胞外表面表达,并且可能与整合素β1具有共同的免疫特性。