Cynomolgus monkey testicular cDNAs for discovery of novel human genes in the human genome sequence
Cynomolgus monkey testicular cDNAs for discovery of novel human genes in the human genome sequence
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DOI：
10.1186/1471-2164-3-36
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发表时间：
2002-01-01
期刊：
影响因子：
4.4
通讯作者：
Hashimoto, K
中科院分区：
文献类型：
--
作者：
Osada, N;Hida, M;Hashimoto, K
背景资料：为了建立完整的人类基因组转录区图谱，我们构建了食蟹猴睾丸cDNA文库，并尝试寻找新的转录本用于鉴定其人类同源物。最终，我们发现302个非冗余cDNA携带300 bp长度或更长的开放阅读框架。其中，89个cDNA在Ensembl人类数据库中未被注释。在检索Ensembl小鼠数据库后，我们还发现69个推定的编码序列在Ensembl中注释的人和小鼠基因组序列中没有同源cDNA。我们随后设计了一个包含396个非冗余cDNA（有和没有开放阅读框）的DNA微阵列，以检查全序列基因的表达。用睾丸探针和其他10种组织的探针混合物，在332个有效点中有316个显示出强杂交信号，75个cDNA在食蟹猴睾丸中显示出非常高的表达，但不是普遍表达。在本报告内，我们确定了302个完整的-插入具有足够长度开放阅读框的食蟹猴cDNA序列，以发现作为人同源物的新转录物。在302个cDNA序列中，89个cDNA的人类同源物在Ensembl的注释的人类基因组序列中没有预测。此外，我们还利用DNA微阵列技术，在全序列克隆中鉴定出75个睾丸中的优势表达基因。我们的cDNA克隆和分析结果将为未来的功能基因组学研究提供宝贵的资源。
Background: In order to contribute to the establishment of a complete map of transcribed regions of the human genome, we constructed a testicular cDNA library for the cynomolgus monkey, and attempted to find novel transcripts for identification of their human homologues.Result: The full-insert sequences of 512 cDNA clones were determined. Ultimately we found 302 non-redundant cDNAs carrying open reading frames of 300 bp-length or longer. Among them, 89 cDNAs were found not to be annotated previously in the Ensembl human database. After searching against the Ensembl mouse database, we also found 69 putative coding sequences have no homologous cDNAs in the annotated human and mouse genome sequences in Ensembl. We subsequently designed a DNA microarray including 396 non-redundant cDNAs (with and without open reading frames) to examine the expression of the full-sequenced genes. With the testicular probe and a mixture of probes of 10 other tissues, 316 of 332 effective spots showed intense hybridized signals and 75 cDNAs were shown to be expressed very highly in the cynomolgus monkey testis, but not ubiquitously.Conclusions: In this report, we determined 302 full-insert sequences of cynomolgus monkey cDNAs with enough length of open reading frames to discover novel transcripts as human homologues. Among 302 cDNA sequences, human homologues of 89 cDNAs have not been predicted in the annotated human genome sequence in the Ensembl. Additionally, we identified 75 dominantly expressed genes in testis among the full-sequenced clones by using a DNA microarray. Our cDNA clones and analytical results will be valuable resources for future functional genomic studies.