EST analysis of mouse retina and RPE/choroid cDNA libraries

[摘要] Purpose: cDNA libraries from the mouse retina have recently beenreported, but no well characterized library from the retinal pigmentepithelium (RPE) or choroid of the mouse has yet appeared in theliterature. To complement these libraries and to provide the first mouseRPE/choroid library, we used freshly dissected tissue from adultC57BL/6J mice to construct new retina and RPE/choroid libraries.Methods: Eyes from 100 six to eight week old C57BL/6J mice weredissected in groups of 10. The whole retina and RPE/choroid wereisolated individually and then homogenized before RNA isolation. Over5000 clones each were sequenced from the unamplified and un-normalizedretina and RPE/choroid libraries. All sequences were analyzed usingGRIST (GRouping and Identification of Sequence Tags), a bioinformaticsprogram for gene identification and clustering.Results: The RPE/choroid library contained 3145 clusters with 76% ofthe clusters representing single clones. Nearly 87% of the clusterscorresponded to named genes in GenBank, and 8% of the RPE clustersremain unidentified. The retina library contained 3190 clusters of which78% represented only one clone. Approximately 85% of the clustersmatched sequences in GenBank, and 9% of the clusters remainunidentified. The clones most abundant in each library were allwell-known sequences and both libraries contained a number of tissuespecific or tissue-enhanced genes.Conclusions: These new libraries should provide a valuable resourcefor gene discovery and cDNAs for expression analysis and functionalstudies.

[学科分类] 生物化学/生物物理