STABLE EXPRESSION OF THE NEURONAL BI (CLASS-A) CALCIUM-CHANNEL IN BABY HAMSTER-KIDNEY CELLS
STABLE EXPRESSION OF THE NEURONAL BI (CLASS-A) CALCIUM-CHANNEL IN BABY HAMSTER-KIDNEY CELLS
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DOI：
10.1006/bbrc.1994.2399
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发表时间：
1994-09-30
影响因子：
3.1
通讯作者：
KATAYAMA, K
中科院分区：
文献类型：
--
作者：
NIIDOME, T;TERAMOTO, T;KATAYAMA, K
将含有二氢叶酸还原酶（DHFR）基因的BI (α (1)) cDNA、含有neo标记基因的骨骼肌α(2)-亚单位cDNA和β -亚单位cDNA的质粒共转染到幼鼠肾（BHK）细胞中。BHK细胞缺乏内源性钙通道活性。利用膜片钳技术，发现20%的甲氨蝶呤（MTX）和G418抗性克隆表达钙通道活性。单克隆BHKBI147在20代传代中表现出稳定的电生理特性。BHKBL147细胞中BI通道的Ca2+电流被一种特定的p型阻滞剂omega-AgaIVA阻断，IC50为150 nM。与BI通道不同，BHK细胞中表达的心脏l型通道Ca2+电流被l型拮抗剂硝苯地平完全阻断，IC50为56 nM。(C) 1994学术出版社，Inc.
Plasmids containing the BI (alpha(1)) cDNA with the dihydrofolate reductase (DHFR) gene, skeletal muscle alpha(2)-subunit cDNA with the neo marker gene, and beta-subunit cDNA were co-transfected into baby hamster kidney (BHK) cells. BHK cells lack endogenous calcium channel activity. Twenty percent of the methotrexate (MTX) and G418 resistant clones were found to express calcium channel activity using the patch-clamp technique. A single clone,BHKBI147, demonstrated stable electrophysiological characteristics over 20 passages. Ca2+ currents of the BI channel in BHKBL147 cells were largely blocked by a specific P-type blocker, omega-AgaIVA, with an IC50 of 150 nM. Unlike the BI channel, Ca2+ currents of cardiac L-type channels expressed in BHK cells were completely blocked by the L-type antagonist, nifedipine, with an IC50 of 56 nM. (C) 1994 Academic Press, Inc.