"neibank: Est Analysis And Bioinformatics For Ocular Gen

眼睛是一个由不同发育来源的高度分化的组织组成的复杂系统。许多对眼睛功能至关重要的基因是组织特异性的，其中许多已知的基因与遗传性眼病有关。大多数人类表达的基因都是通过表达序列标签(EST)才知道的。然而，到目前为止，许多眼组织在参与DBEST的cDNA文库中的代表性很差。为了解决这种情况，已经启动了一个名为NEIBank的项目，以提高人类眼睛的EST覆盖率，并为眼睛开发一部分子百科全书。构建了人晶状体、睫状体/虹膜、RPE/脉络膜、视网膜和角膜的cDNA文库，并进行了深度测序。来自每个非标准化文库的2,000到10,000个克隆已经使用定制软件GRIST进行了测序、分析和聚类。镜头和虹膜资料库已被标准化，以获得更深层次的序列。还建立了大鼠全眼、虹膜/睫状体/TM、视网膜和小鼠全眼文库。许多新的基因已经被发现，包括opticin/oculoglycan(一种眼睛特异的富含亮氨酸的重复蛋白，其基因图谱接近ARMD)，Lgs(一种丰富的晶状体特异性转录本，与谷氨酰胺合成酶和念珠菌晶体蛋白脱酰胺酶相关)，oculospanin(一种新的四聚糖膜蛋白，其基因定位于chr17，靠近疾病基因点RP17)，虹膜表达的生长因子(与血管内皮生长因子相关)和retbindin，一种与氟立酮结合蛋白相关的新的视网膜分泌蛋白。此外，我们已经确定了许多重要的眼睛转录本的选择性剪接形式，这些转录本可以产生修饰的蛋白质。这些包括更大版本的晶状体膜蛋白MP19/Lim2和修饰版本的NRL，视网膜转录因子，以及视网膜色素上皮的蛋白质Bestrophin。总共有大约20,000个眼部基因克隆，代表了超过12,000个单独的GRIST簇(潜在基因)，已经被测序并组织在一个包含关键字和染色体位置数据的基于网络的数据库(neibank.nei.nih.gov)中。这项工作产生的DNA资源现在正被用于创建眼部研究的微阵列。GRIST程序也被用于分析NIDCD项目的类似数据。

The eye is a complex system of highly differentiated tissues of various developmental origins. Many genes essential for eye function are tissue-specific and many of those known are associated with genetic eye diseases. The majority of human expressed genes are known only through expressed sequence tags (ESTs). However many eye tissues have been poorly represented in the cDNA libraries that have so far contributed to dbEST. To address this situation a project, NEIBank, has been initiated to improve the EST coverage of the human eye and to develop a molecular encyclopedia for the eye. cDNA libraries for human lens, ciliary body/iris,RPE/choroid, retina and cornea have been constructed and examined by in-depth sequencing. Between 2,000 and 10,000 clones from each un-normalized library have been sequenced, analyzed and clustered using custom software, GRIST. Lens and iris libraries have been normalized for deeper sequence. Libraries for rat whole eye, iris/ciliary body/TM and retina and mouse whole eye have also been created. Many novel genes have been identified, including opticin/oculoglycan (an eye-specific leucine-rich repeat protein whose gene maps close to ARMD), lengsin (LGS)(an abundant lens specific transcript related to glutamine synthetase and a candiate crystallin deamidase), oculospanin (a novel tetraspan membrane protein whose gene maps to chr17, close to the disease locus RP17), iris-expressed growth factor (a VEGF relative) and retbindin, a novel secreted protein of the retina that is related to flavionoid binding proteins. In addition we have identifed numerous alternative splice forms of important eye transcripts that could give rise to modified proteins. These include a larger version of the lens membrane protein MP19/Lim2 and modifed versions of Nrl, a retina transcriptoin factor, and Bestrophin, a protein of the retinal pigment epithelium. In total, approximately 20,000 ocular cDNA clones, representing over 12,000 individual GRIST clusters (potential genes), have been sequenced and organized in a web-based database (neibank.nei.nih.gov)that contains keyword and chromosomal location data. The DNA resource generated by this work is now being used to create micro-arrays for ocular studies. The GRIST procedure is also being used to analyse similar data for projects in NIDCD.