A RecA-mediated exon profiling method

Yuki Hasegawa, Shiro Fukuda, Kazuro Shimokawa, Shinji Kondo, Norihiro Maeda, Yoshihide Hayashizaki

研究成果: Article

4 被引用数 (Scopus)

抄録

We have developed a RecA-mediated simple, rapid and scalable method for identifying novel alternatively spliced full-length cDNA candidates. This method is based on the principle that RecA proteins allow to carry radioisotope-labeled probe DNAs to their homologous sequences, resulting in forming triplexes. The resulting complex is easily detected by mobility difference on electrophoresis. We applied this exon profiling method to four selected mouse genes as a feasibility study. To design probes for detection, the information on known exonic regions was extracted from public database, RefSeq. Concerning the potentially transcribed novel exonic regions, RNA mapping experiment using Affymetrix tiling array was performed. As a result, we were able to identify alternative splice variants of Thioredoxin domain containing 5, Interleukin1β, Interleukin 1 family 6 and glutamine-rich hypothetical protein. In addition, full-length sequencing demonstrated that our method could profile exon structures with >90% accuracy. This reliable method can allow us to screen novel splice variants from a huge number of cDNA clone set effectively.

本文言語English
論文番号e97
ジャーナルNucleic acids research
34
13
DOI
出版ステータスPublished - 2006 9 22
外部発表はい

ASJC Scopus subject areas

  • Genetics

フィンガープリント 「A RecA-mediated exon profiling method」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。

引用スタイル