Maruyama Yukio, Wakamatsu Ai, Kawamura Yoshifumi, Kimura Kouichi, Yamamoto Jun-ichi, Nishikawa Tetsuo, Kisu Yasutomo, Sugano Sumio, Goshima Naoki, Isogai Takao, Nomura Nobuo
Japan Biological Informatics Consortium, Aomi, Koto-ku, Tokyo 135-8073, Japan.
Nucleic Acids Res. 2009 Jan;37(Database issue):D762-6. doi: 10.1093/nar/gkn872.
Completion of human genome sequencing has greatly accelerated functional genomic research. Full-length cDNA clones are essential experimental tools for functional analysis of human genes. In one of the projects of the New Energy and Industrial Technology Development Organization (NEDO) in Japan, the full-length human cDNA sequencing project (FLJ project), nucleotide sequences of approximately 30 000 human cDNA clones have been analyzed. The Gateway system is a versatile framework to construct a variety of expression clones for various experiments. We have constructed 33 275 human Gateway entry clones from full-length cDNAs, representing to our knowledge the largest collection in the world. Utilizing these clones with a highly efficient cell-free protein synthesis system based on wheat germ extract, we have systematically and comprehensively produced and analyzed human proteins in vitro. Sequence information for both amino acids and nucleotides of open reading frames of cDNAs cloned into Gateway entry clones and in vitro expression data using those clones can be retrieved from the Human Gene and Protein Database (HGPD, http://www.HGPD.jp). HGPD is a unique database that stores the information of a set of human Gateway entry clones and protein expression data and helps the user to search the Gateway entry clones.
人类基因组测序的完成极大地加速了功能基因组学研究。全长cDNA克隆是人类基因功能分析必不可少的实验工具。在日本新能源和工业技术开发组织(NEDO)的一个项目——全长人类cDNA测序项目(FLJ项目)中,已经分析了约30000个人类cDNA克隆的核苷酸序列。Gateway系统是一个通用框架,可用于构建各种用于不同实验的表达克隆。我们从全长cDNA构建了33275个人类Gateway入门克隆,据我们所知,这是世界上最大的此类克隆集合。利用这些克隆以及基于小麦胚芽提取物的高效无细胞蛋白质合成系统,我们已经在体外系统地、全面地生产并分析了人类蛋白质。克隆到Gateway入门克隆中的cDNA开放阅读框的氨基酸和核苷酸序列信息以及使用这些克隆的体外表达数据可从人类基因与蛋白质数据库(HGPD,http://www.HGPD.jp)中检索。HGPD是一个独特的数据库,它存储了一组人类Gateway入门克隆的信息和蛋白质表达数据,并帮助用户搜索Gateway入门克隆。