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从一个富集的成人视网膜文库中鉴定出的cDNA克隆的表达谱和染色体定位。

Expression profile and chromosomal location of cDNA clones, identified from an enriched adult retina library.

作者信息

Sinha S, Sharma A, Agarwal N, Swaroop A, Yang-Feng T L

机构信息

Department of Genetics, Yale University School of Medicine, New Haven, CT, USA.

出版信息

Invest Ophthalmol Vis Sci. 2000 Jan;41(1):24-8.

Abstract

PURPOSE

To delineate the profile of genes expressed in the adult human retina and assign chromosomal location of cDNA clones.

METHODS

The end-sequence of random clones from an enriched human retinal cDNA library was analyzed by NCBI database search. Expression profile was established by northern blot analysis, database search, or both. Selected cDNA clones were localized to human chromosomes by somatic cell hybrid analysis, in situ hybridization to metaphase chromosomes, or both. Chromosomal location was also obtained by searching the databases.

RESULT

One hundred and thirty-seven clones were isolated from the subtracted retinal library. Fifty-one clones were identical with 35 known human genes in GenBank, and 24 clones corresponded to 23 uncharacterized human expressed sequenced tags (ESTs), novel genes, or both. The remaining 59 clones were not pursued further because they contained bacterial sequences or repetitive elements. Several clones indicated a restricted pattern of expression with high levels of transcripts in the retina. Chromosomal location of novel retinal ESTs is also reported.

CONCLUSIONS

This study provides a profile of genes expressed in the adult human retina. One round of subtraction eliminated most constitutively expressed genes and permitted partial normalization of the retinal library. Twenty-three novel genes were identified. The combined information obtained from expression analysis and chromosomal localization of retinal cDNAs should be valuable in identifying candidate genes for diseases involving retinal dysfunction.

摘要

目的

描绘成人人类视网膜中表达的基因图谱,并确定cDNA克隆的染色体定位。

方法

通过NCBI数据库搜索分析来自富集的人类视网膜cDNA文库的随机克隆的末端序列。通过Northern印迹分析、数据库搜索或两者来建立表达谱。通过体细胞杂交分析、中期染色体原位杂交或两者将选定的cDNA克隆定位到人类染色体上。染色体定位也通过搜索数据库获得。

结果

从消减视网膜文库中分离出137个克隆。51个克隆与GenBank中35个已知人类基因相同,24个克隆对应于23个未表征的人类表达序列标签(EST)、新基因或两者。其余59个克隆未进一步研究,因为它们包含细菌序列或重复元件。几个克隆显示出在视网膜中转录本水平较高的受限表达模式。还报道了新的视网膜EST的染色体定位。

结论

本研究提供了成人人类视网膜中表达的基因图谱。一轮消减消除了大多数组成性表达的基因,并使视网膜文库部分标准化。鉴定出23个新基因。从视网膜cDNA的表达分析和染色体定位获得的综合信息对于鉴定涉及视网膜功能障碍疾病的候选基因应该是有价值的。

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