The status, quality, and expansion of the NIH full-length cDNA project: The Mammalian Gene Collection (MGC)
The status, quality, and expansion of the NIH full-length cDNA project: The Mammalian Gene Collection (MGC)
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DOI：
10.1101/gr.2596504
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发表时间：
2004-10-01
期刊：
影响因子：
7
通讯作者：
Collins, FS
中科院分区：
文献类型：
--
作者：
Gerhard, DS;Wagner, L;Collins, FS
美国国立卫生研究院的哺乳动物基因保藏中心 (MGC) 项目旨在生成并测序可公开访问的 cDNA 资源，其中包含每个人类和小鼠基因的完整开放阅读框 (ORF)。该项目最初使用随机策略从来自不同组织的大量 cDNA 文库中选择克隆。根据 5'-EST 序列选择候选克隆，然后进行高精度的完整测序，并通过为本项目开发的算法进行分析。目前，MGC 中至少有一个具有完整 ORF 的克隆代表了超过 11,000 个人类基因和 10,000 个小鼠基因。随机选择方法现已达到饱和点，现在需要过渡到针对缺失转录本的方案以完成小鼠和人类的收集。 MGC 克隆的序列与参考基因组序列的比较表明，大多数 cDNA 克隆具有非常高的序列质量，尽管某些 cDNA 可能存在！由于实验假象（例如 PCR、克隆或逆转录酶错误），可能会携带错义变异。最近，大鼠 cDNA 组件被添加到该项目中，并且正在进行的青蛙 (Xenopus) 和斑马鱼 (Danio) cDNA 项目也得到了扩展，以利用高通量 MGC 管道。
The National Institutes of Health's Mammalian Gene Collection (MGC) project was designed to generate and sequence a publicly accessible cDNA resource containing a complete open reading frame (ORF) for every human and mouse gene. The project initially used a random strategy to select clones from a large number of cDNA libraries from diverse tissues. Candidate clones were chosen based on 5'-EST sequences, and then fully sequenced to high accuracy and analyzed by algorithms developed for this project. Currently, more than 11,000 human and 10,000 mouse genes are represented in MGC by at least one clone with a full ORF. The random selection approach is now reaching a saturation point, and a transition to protocols targeted at the missing transcripts is now required to complete the mouse and human collections. Comparison of the sequence of the MGC clones to reference genome sequences reveals that most cDNA clones are of very high sequence quality, although it is likely that some cDNA! may carry missense variants as a consequence of experimental artifact, such as PCR, cloning, or reverse transcriptase errors. Recently, a rat cDNA component was added to the project, and ongoing frog (Xenopus) and zebrafish (Danio; cDNA projects were expanded to take advantage of the high-throughput MGC pipeline.