Keyue Ding, Xiaojing Sun, Min Zhou, Jing Cai, Tianjing Cai, Yan Zhang, Tongbin Zhu, Zhengguo Zhang, B. Qiang, Yan Shen
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引用次数: 2
摘要
我们提出了一种从我们以前的研究中产生的人类胎儿海马ESTs数据库中鉴定新的蛋白质编码基因的方法。该协议涉及到对GenBank非冗余(nr)蛋白数据库进行ESTs编码区域查询。利用ESTScan程序对ESTs序列数据中的编码区域进行检测和重构。在分析了近10,000个原始ESTs后,提取了4,061个编码区域。然后在GenBank nr-protein数据库中检索相应的氨基酸序列;所有相似点都在同源性分析工作台中进行解析。22条est被鉴定为“种子”est,可能是新蛋白编码基因的片段。事实上,通过对这些est的芯片全长cDNA克隆的进一步分析,以及UCSC中的定位信息和增加的实验证明,验证了该策略的有效性。利用稀缺的est资源和有效的策略,我们将在人类基因组中探索更多的新基因。[本文中描述的ESTs序列数据已提交至GenBank数据库(http://www.ncbi.nlm.nih.gov/dbEST), accession no . BM174886-958, BM259607-9;全长cDNA序列可从http://www.chgb.org.cn/hippocampus.htm获取]
Identification of novel protein-coding genes from human fetal hippocampus ESTs database
We presented a procedure to identify novel protein-coding genes from human fetal hippocampus ESTs database generated in our previous study. The protocol is involved in searching the GenBank non-redundant (nr) protein database with ESTs coding regions query. We used ESTScan program to detect and reconstruct coding regions in the ESTs sequence data. With almost 10,000 raw ESTs analyzed, 4,061 coding regions were extracted. The corresponding amino acid sequences were then searched against the GenBank nr-protein databases; all the similarity hits were parsed in a homology analysis workbench. 22 ESTs were identified as the “seed” ESTs, which might be the fragment of the novel protein-coding genes. Indeed, further analysis of these ESTs in full-length cDNA cloning in silico, as well as the mapping information in UCSC and the added experimental proof, verified the effectiveness of the strategy. Taking advantage of rare ESTs resources, and the effective strategy, we would explore more novel genes in the human genome.
[The ESTs sequence data described in this paper have been submitted to the GenBank data library (http://www.ncbi.nlm.nih.gov/dbEST) under accession nos. BM174886-958, BM259607-9; and the full length cDNA sequence can be obtained in http://www.chgb.org.cn/hippocampus.htm]