Stocker M, Kerschensteiner D
Max-Planck Institut für Experimentelle Medizin, Göttingen, Germany.
Biochem Biophys Res Commun. 1998 Jul 30;248(3):927-34. doi: 10.1006/bbrc.1998.9072.
The expressed sequence tag (EST) database is a valuable tool to identify functionally related clones, when sequence similarity is so low that standard library screening methods cannot be successfully applied. Comparing conserved protein sequences of cloned voltage-gated potassium channels led to the identification and cloning of a new putative potassium channel alpha-subunit from rat brain, Kv9.1. A homologue of this alpha-subunit was also found in human brain tissue. A second alpha-subunit, Kv9.3, most probably belonging to the same subfamily, was also isolated and sequenced. Tissue distribution and analysis of genomic DNA were performed for both channels. rKv9.1 is expressed exclusively in the central nervous system, whereas rKv9.3 shows a widespread tissue distribution. No currents could be measured in X. oocytes upon injection of rKv9.1 or rKv9.3 cRNA.
当序列相似性非常低以至于无法成功应用标准文库筛选方法时,表达序列标签(EST)数据库是鉴定功能相关克隆的宝贵工具。通过比较克隆的电压门控钾通道的保守蛋白序列,从大鼠脑中鉴定并克隆出一种新的假定钾通道α亚基Kv9.1。在人类脑组织中也发现了该α亚基的同源物。还分离并测序了第二个α亚基Kv9.3,它很可能属于同一亚家族。对这两种通道都进行了组织分布和基因组DNA分析。rKv9.1仅在中枢神经系统中表达,而rKv9.3则表现出广泛的组织分布。注射rKv9.1或rKv9.3的cRNA后,在非洲爪蟾卵母细胞中未检测到电流。