Purpose. To clone and characterize the mammalian homolog of the Drosophila ring canal protein kelch (mRCP) and determine its role in the development of the RPE and in RPE/retina interaction, kelch is involved in regulating cytoplasmic transport between nurse cells and the oocyte during oogenesis. The existence of a RCP homolog was first suggested by sequencing of clones from a subtracted library enriched for cDNAs expressed preferentially in the RPE and retina. Methods. Subtractive hybridization and random sequencing were used to isolate the clone from a bovine eye cup cDNA library. The subtractive library was generated as described (Chang, et.al., ARVO 1996). RT-PCR, library screening, in situ hybridization and DNA sequencing are being performed by standard methods. Results. The 237bp mRCP clone shows 57% homology to ORF2 of the 5.6kb Drosophila kelch gene. The blastx probability score (4. le-21) is highly significant. In addition, the mRCP shows homology to ESTs cloned from human retina, liver and fetal brain. Efforts are currently underway to obtain a full length cDNA and to determine the spatial and developmental expression pattern of mRCP. Conclusions The results suggest that a mRCP homolog exists and that it is expressed in the retina and/or RPE. Based on the function of kelch in Drosophila, mRCP may be involved in retina/RPE interaction during development.
|Original language||English (US)|
|Journal||Investigative Ophthalmology and Visual Science|
|State||Published - Feb 15 1996|
ASJC Scopus subject areas
- Sensory Systems
- Cellular and Molecular Neuroscience